Publications (92)285.6 Total impact
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Article: Molecular cloning, phylogenetic analysis and functional characterization of soluble Toll-like receptor 5 in gilthead seabream, Sparus aurata.
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ABSTRACT: Two forms of TLR5, one membrane-anchored and one soluble, have been described in some teleost fish species. However, the exact role of each form has been little studied. In the present study, we show that the mRNA levels of soluble gilthead seabream TLR5 (sbTLR5S) are highly induced in head kidney, spleen, liver and blood after Vibrio anguillarum infection, suggesting an important role for sbTLR5S in the innate immune response against bacteria. Comparative genomic and phylogenetic analyses revealed a co-evolution pattern of both genes across fish species and a proximal location in their genomes, further suggesting a functional link between them. To further investigate this issue, the coding sequence of the sbTLR5S was cloned and the corresponding recombinant protein was produced in HEK cells. The gene product was secreted to the culture medium as a soluble factor and a physical interaction between flagellin and sbTLR5S was demonstrated. Collectively, these results suggest that sbTLR5S plays an important role in modulating the flagellin-mediated immune response in seabream.Fish & Shellfish Immunology 04/2013; · 3.32 Impact Factor -
Dataset: Galindo-Villegas - PNAS 2012
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Article: Regulation of natural killer enhancing factor (NKEF) genes in teleost fish, gilthead seabream and European sea bass.
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ABSTRACT: Peroxiredoxins (Prx) are a family of antioxidant proteins also involved in inflammation and innate immunity. Prx1 and Prx2 are also known as natural killer enhancing factor (NKEF)-A and NKEF-B, respectively, by their ability to prime the mammalian NK-cells activity. In teleost fish, NKEF genes have been isolated but their regulation has been scarcely evaluated. We have identified orthologues of the NKEF-A and NKEF-B genes in the teleost European sea bass (Dicentrarchus labrax) which showed constitutive expression and wide distribution in their tissues. In vitro, the gilthead seabream (Sparus aurata) and sea bass NKEFs were slightly up-regulated in head-kidney leucocytes after stimulation with unmethylated CpG oligodeoxynucleotides, poly I:C or pathogenic bacteria. In vivo, seabream and sea bass infection with nodavirus up-regulated the expression of NKEF genes in the brain (target tissue for nodavirus) and head-kidney at different infection times. Although further studies are necessary to ascertain their role as antioxidant proteins and in the immune response in teleost fish, our results suggest a primary role of seabream and sea bass NKEFs in the innate immune response against bacterial and viral agents.Molecular Immunology 03/2013; · 2.90 Impact Factor -
Article: Changes in intestinal morphology and microbiota caused by dietary administration of inulin and Bacillus subtilis in gilthead sea bream (Sparus aurata L.) specimens.
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ABSTRACT: Changes produced in gilthead sea bream (Sparus aurata L.) intestinal morphology and microbiota caused by dietary administration of inulin and Bacillus subtilis have been studied. Gilthead sea bream specimens were fed diets containing 0 (control), inulin (10 g kg(-1)), B. subtilis (10(7) cfu g(-1)), or B. subtilis + inulin (10(7) cfu g(-1) + 10 g kg(-1)) for four weeks. Curiously, fish fed the experimental diets (inulin, B. subtilis, or B. subtilis + inulin) showed the same morphological alterations when studied by light and electron microscopy, while significant differences in the signs of intestinal damage were detected by the morphometric study. All of the observed alterations were present only in the gut mucosa, and intestinal morphometric study revealed no effect of inulin or B. subtilis on the intestinal absorptive area. Furthermore, experimental diets cause important alterations in the intestinal microbiota by significantly decreasing bacterial diversity, as demonstrated by the specific richness, Shannon, and range-weighted richness indices. The observed alterations demonstrate that fish fed experimental diets had different signs of gut oedema and inflammation that could compromise their body homeostasis, which is mainly maintained by the epithelial lining of the gastrointestinal tract. To our knowledge, this is the first in vivo study regarding the implications of the use of synbiotics (conjunction of probiotics and prebiotics) on fish gut morphology and microbiota.Fish & Shellfish Immunology 02/2013; · 3.32 Impact Factor -
Article: Effects of dietary inulin, B. subtilis and microalgae on intestinal gene expression in gilthead seabream (Sparus aurata L.).
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ABSTRACT: The present work describes effects of dietary inulin, two microalgae (Tetraselmis chuii and Phaeodactylum tricornutum) and Bacillus subtilis (solely or combined with insulin or microalgae) on the expression of different genes in the intestine of the gilthead seabream (Sparus aurata L.) following four weeks of a feeding trial. Selected genes were grouped into five categories: genes involved in inflammation (genes encoding proinflammatory proteins), genes related to the cytoskeleton, genes encoding proteins of junction complexes, genes implicated in digestion processes and genes related to transport proteins. Regarding proinflammatory genes, interleukin-8 (IL-8) expression showed a significant increase in the fish fed all the assayed diets, except the B. subtilis + inulin diet, whereas the expression of caspase-1 (CASP-1) was also increased by the B. subtilis and B. subtilis + T. chuii diets. Cyclooxygenase-2 (COX-2) gene expression only increased in fish fed the B. subtilis diet. Among cytoskeletal and junctional genes, only β-actin and occludin were significantly affected by the assayed diets. β-actin expression was up-regulated by the inulin-containing diets (inulin and B. subtilis + inulin diets), whereas occludin expression increased in the fish fed all the assayed diets, except the P. tricornutum diet. Finally, the expression of transport protein genes demonstrated that the inulin diet and all the experimental diets containing B. subtilis significantly increased transferrin expression, whereas digestive enzymes were not affected by the experimental diets. In conclusion, our results demonstrated that inulin, B. subtilis and microalgae can modulate intestinal gene expression in the gilthead seabream. To our knowledge, this is the first study of the effects of some food additives on the intestinal expression of different genes in this species. More studies are needed to understand the role of these genes in maintaining the integrity and functionality of the intestine.Fish & Shellfish Immunology 01/2013; · 3.32 Impact Factor -
Article: Influence of melatonin on the immune system of fish: a review.
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ABSTRACT: Endocrine-immune system interactions have been widely demonstrated in mammals, whereas in fish, these relationships remain unclear. Of the organs that constitute the endocrine system, the pineal gland and its secretory product melatonin act in the synchronization of daily and seasonal rhythms in most vertebrates, including fish. Seasonal differences in immunocompetence and disease prevalence have been well documented in humans. Seasonality also strongly influences the life history of fish by controlling the timing of physiological events, such as reproduction, food intake, locomotor activity, and growth performance. Apart from its synchronizing capabilities, the role of melatonin in physiological processes in fish is not thoroughly understood. The purpose of this review is to summarize current studies on the effects of melatonin on the fish immune system. These studies suggest that melatonin represents an important component of fish endocrine-immune system interactions. The elucidation of the defense mechanisms of fish will facilitate the development of health management tools to support the growing finfish aquaculture industry as well as address questions concerning the origins and evolution of the immune system in vertebrates.International Journal of Molecular Sciences 01/2013; 14(4):7979-99. · 2.60 Impact Factor -
Article: Histological alterations and microbial ecology of the intestine in gilthead seabream (Sparus aurata L.) fed dietary probiotics and microalgae.
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ABSTRACT: The effects on histology and microbial ecology in gilthead seabream (Sparus aurata) intestine caused by dietary probiotic and microalgae were studied. Fish were fed non-supplemented (C, control) or supplemented diets with Tetraselmis chuii, Phaeodactylum tricornutum and Bacillus subtilis single or combined (diets T, P, B, BT and BP) for 4 weeks. Curiously, fish fed the experimental diets showed similar morphological alterations when studied by light and electron microscopy and significant signs of intestinal damage were detected. No effect of microalgae or B. subtilis on the intestinal absorptive area was observed, whereas the number of goblet cells and IELs were significantly lower in fish fed the T, P, B and BT diets and T, BT and BP diets, respectively. Interestingly, only the diets containing B. subtilis resulted in a significant reduction of microvilli height. Alterations such as wide intercellular spaces and large vacuoles in enterocytes were observed in fish fed T, B, BT, BT and P in lesser degrees. These observations demonstrate that fish fed experimental diets presented different signs of oedema and inflammation that could compromise their body homeostasis. Moreover, the experimental diets cause important alterations in the intestinal microbiota by a significant decrease in bacterial diversity, as demonstrated by the fall in specific richness, Shannon and range-weighted richness indices. To our knowledge, this is the first in vivo study regarding the implications of the use of probiotics in combination with immunostimulants on fish intestinal morphology and microbiota. More morphofunctional studies are needed in order to correlate the nutritional and immune aspects of fish gut.Cell and Tissue Research 10/2012; · 3.11 Impact Factor -
Article: Nodavirus infection induces a great innate cell-mediated cytotoxic activity in resistant, gilthead seabream, and susceptible, European sea bass, teleost fish.
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ABSTRACT: Viral nervous necrosis (VNN) virus produces great mortalities in fish having susceptible and reservoir species between the most important marine aquaculture species. Cell-mediated cytotoxicity (CMC) is considered, towards the interferon (IFN), the most important mechanism of the immune response to fight against viral infections but it has been very scarcely evaluated. We aimed to evaluate the effects of VNNV infection in the reservoir gilthead seabream (Sparus aurata) and susceptible European sea bass (Dicentrarchus labrax). Firstly, after experimental infection we found mortalities in the sea bass (55%) but no in the seabream. Moreover, VNN virus replicates in the brain of both species as it was reflected by the high up-regulation of the Mx gene expression. Interestingly, the head-kidney leucocyte cell-mediated cytotoxic activity was significantly increased in both species reaching highest activity at 7 days: 3.65- and 2.7-fold increase in seabream and sea bass, respectively. This is supported by the significant up-regulation of the non-specific cytotoxic cell receptor (NCCRP-1) in the two fish species. By contrast, phagocytosis was unaffected in both species. The respiratory burst was increased in seabream 7 days post-infection whilst in sea bass this activity was significantly decreased at days 7 and 15. Our results demonstrate the significance of the CMC activity in both gilthead seabream and European sea bass against nodavirus infections but further studies are still needed to understand the role of cytotoxic cells in the antiviral immune response and the mechanisms involved in either reservoir or susceptible fish species.Fish & Shellfish Immunology 09/2012; 33(5):1159-66. · 3.32 Impact Factor -
Article: TNF receptors regulate vascular homeostasis through a caspase-8, caspase-2 and P53 apoptotic program that bypasses caspase-3.
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ABSTRACT: Although it is known that TNF receptor signaling plays a critical role in vascular integrity and homeostasis, the contribution of each receptor to these processes and the signaling pathway involved are still largely unknown. Here, we show that targeted gene knockdown of TNFRSF1B in zebrafish embryos results in the induction of a caspase-8, caspase-2 and P53-dependent apoptotic program in endothelial cells that bypasses caspase-3. Furthermore, the simultaneous depletion of TNFRSF1A or the activation of NF-κB rescue endothelial cell apoptosis, indicating that a signaling balance between both TNFRs is required for endothelial cell integrity and that, in endothelial cells, TNFRSF1A signals apoptosis through caspase-8, while TNFRSF1B signals survival via NF-κB. Similarly, TNFα promotes the apoptosis of human endothelial cells through TNFRSF1A and triggers caspase-2 and P53 activation. We have identified an evolutionary conserved apoptotic pathway involved in vascular homeostasis that provides new therapeutic targets for the control of inflammation and tumor driven angiogenesis.Disease Models and Mechanisms 09/2012; · 4.94 Impact Factor -
Article: Regulation of immunity and disease resistance by commensal microbes and chromatin modifications during zebrafish development.
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ABSTRACT: How fish larvae are protected from infection before the maturation of adaptive immunity, a process which may take up to several weeks in most species, has long been a matter of speculation. Using a germ-free model, we show that colonization by commensals in newly hatched zebrafish primes neutrophils and induces several genes encoding proinflammatory and antiviral mediators, increasing the resistance of larvae to viral infection. Commensal microbe recognition was found to be mediated mainly through a TLR/MyD88 signaling pathway, and professional phagocytes were identified as the source of these immune mediators. However, the induction of proinflammatory and antiviral genes, but not of antimicrobial effector genes, also required the covalent modification of histone H3 at gene promoters. Interestingly, chromatin modifications were not altered by commensal microbes or hatching. Taken together, our results demonstrate that gene-specific chromatin modifications are associated with the protection of zebrafish larvae against infectious agents before adaptive immunity has developed and prevent pathologies associated with excessive inflammation during development.Proceedings of the National Academy of Sciences 09/2012; 109(39):E2605-14. · 9.68 Impact Factor -
Article: Enrichment of gilthead seabream (Sparus aurata L.) diet with microalgae: effects on the immune system.
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ABSTRACT: The present study assessed the effects of three orally administered microalgae (Nannochloropsis gaditana, Tetraselmis chuii and Phaeodactylum tricornutum) on different immune parameters and immune-related gene expression of gilthead seabream (Sparus aurata L.). Fish were fed a control or one of six experimental diets and sampled at 2 and 4 weeks of treatment. At the end of the trial, growth performance and different systemic and local immune activities were measured (natural haemolytic complement activity, IgM levels, phagocytosis and respiratory burst). The expression levels of different immune-associated genes (EF-1α, IgM(H), TCR-β, MHCIα, MHCIIα, CSF-1R and β-defensin) were analysed in head-kidney and gut. Administration of N. gaditana and T. chuii resulted in a significant increase in haemolytic complement activity, phagocytic capacity, as well expression level of β-defensin, as well as MHCIIα and CSF-1R, respectively. The P. tricornutum-supplemented diet provoked immunostimulation, and very little effect on gene expression was observed. These results suggest that dietary microalgae enhance gilthead seabream defence activity, which could be very important in order to consider such microalgae as a possible additive in fish diets.Fish Physiology and Biochemistry 06/2012; · 1.53 Impact Factor -
Article: Increases in immune parameters by inulin and Bacillus subtilis dietary administration to gilthead seabream (Sparus aurata L.) did not correlate with disease resistance to Photobacterium damselae.
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ABSTRACT: The present work evaluates the effects of inulin and Bacillus subtilis, single or combined, on immune parameters, immune-related gene expression and protection against Photobacterium damselae subsp. piscicida in gilthead seabream (Sparus aurata). Three trials were conducted. In the first trial, different concentrations of inulin (10, 15 and 30 g kg(-1)) (as a prebiotic) were administered to determine the optimal concentration for stimulating the seabream's immune system. In the second trial, the optimum concentration of inulin (10 g kg(-1)) was combined with B. subtilis (as a probiotic). Following two and four weeks of the treatment, the main immune parameters, as well as the expression of seven immune-related genes, were measured. In the final trial, fish fed the same diet as in the second trial were challenged intraperitoneally with P. damselae subsp. piscicida (10(9) cfu g(-1)). Treatment groups for the second and third trial were control (non-supplemented diet), inulin (10 g kg(-1)), B. subtilis (10(7) cfu g(-1)) and inulin + B. subtilis (10 g kg(-1) and 10(7) cfu g(-1) respectively). Dietary administration of inulin or B. subtilis for two weeks stimulated the serum complement activity and the IgM level, as well as leucocyte phagocytic activity; furthermore, inulin stimulated leucocyte respiratory burst activity. When inulin and B. subtilis were administered together (as a synbiotic), only the serum complement activity and the IgM level increased in a statistically significant manner. Furthermore, the complement activity showed a significant increase in fish fed the three experimental diets for four weeks. The challenge experiment showed that the fish fed inulin or the synbiotic diet had non-significantly lower or significantly higher cumulative mortality, respectively, compared with the control group (non-supplemented diet). These results suggest that inulin and B. subtilis modulate the immune response of the gilthead seabream, although the combined administration increases susceptibility to infection by P. damselae subsp. piscicida.Fish & Shellfish Immunology 06/2012; 32(6):1032-40. · 3.32 Impact Factor -
Article: Effects of dietary Bacillus subtilis, Tetraselmis chuii, and Phaeodactylum tricornutum, singularly or in combination, on the immune response and disease resistance of sea bream (Sparus aurata L.).
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ABSTRACT: Combined or individual effects of two microalgae (Phaeodactylum tricornutum and Tetraselmis chuii) and Bacillus subtilis on immune response, gene expression, and survival to challenge with Photobacterium damselae subsp. piscicida of gilthead sea bream were investigated. To test the capacity of B. subtilis to grow employing the microalgae polysaccharides as energy and carbon source, an in vitro assay was defined, and demonstrated that the digestion product of microalgae, mainly P. tricornutum, supported the growth of B. subtilis much better than glucose. For the in vivo study, fish were distributed in six equal groups (each of two replicates) and received one of the following experimental diets: C) control, non-supplemented diet; T) T. chuii 100 g kg(-1); P) P. tricornutum 100 g kg(-1); B) B. subtilis (10(7) cfu g(-1)); BT) B. subtilis (10(7) cfu g(-1))+T. chuii (100 g kg(-1)); and BP) B. subtilis (10(7) cfu g(-1))+P. tricornutum (100 g kg(-1)). The complement activity, serum IgM level, respiratory burst, phagocytic activity, and expression of seven selected immune-related genes in head-kidney were evaluated following two and four weeks of treatment. At the end of the feeding trial, fish were challenged by intraperitoneal injection of LD(50) concentration of P. damselae subsp. piscicida and mortality was recorded. This is the first study testing the immunomodulatory capacity of the microalgae used in the present work. The dietary applications of B. subtilis, T. chuii, and P. tricornutum, singly or in combination, may exhibit up-regulating effects on gilthead sea bream immune parameters. P. tricornutum demonstrated the highest immunostimulant activity. There were no significant differences between combination feeding and feeding ingredients separately. Our results demonstrated the potential of microalgae as immunostimulants for fish, although further studies regarding the implications and effects of a stimulated immune system against pathogens, especially the protective capacity against specific diseases, are necessary.Fish & Shellfish Immunology 05/2012; 33(2):342-9. · 3.32 Impact Factor -
Article: Evolution of inflammasome functions in vertebrates: Inflammasome and caspase-1 trigger fish macrophage cell death but are dispensable for the processing of IL-1β
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ABSTRACT: Members of the nucleotide binding and oligomerization domain-like receptors (NLRs) and the PYD and CARD domain containing adaptor protein (PYCARD) assemble into multi-protein platforms, termed inflammasomes, to mediate in the activation of caspase-1 and the subsequent secretion of IL-1β and IL-18, and the induction of pyroptotic cell death. While the recognition site for caspase-1 is well conserved in mammals, most of the non-mammalian IL-1β genes cloned so far lack this conserved site. We report here that stimulation or infection of seabream macrophages (MØ) led to the caspase-1-independent processing and release of IL-1β. In addition, several classical activators of the NLRP3 inflammasome failed to activate caspase-1 and to induce the processing and release of IL-1β. Furthermore, the processing of IL-1β in seabream MØ is not prevented by caspase-1 or pan-caspase inhibitors, and recombinant seabream caspase-1 failed to process IL-1β. However, the pharmacological inhibition of caspase-1 impaired Salmonella enterica sv. Typhimurium-induced cell death. These results suggest a role for the inflammasome and caspase-1 in the regulation of pyroptotic cell death in fish and support the idea that its use as a molecular platform for the processing of pro-inflammatory cytokines arose after the divergence of fish and tetrapods.Innate Immunity 03/2012; · 4.00 Impact Factor -
Article: Viral nervous necrosis virus persistently replicates in the central nervous system of asymptomatic gilthead seabream and promotes a transient inflammatory response followed by the infiltration of IgM+ B lymphocytes.
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ABSTRACT: The viral nervous necrosis virus (VNNV) is the causal agent of viral encephalopathy and retinopathy (VER), a worldwide fish disease that is responsible for high mortality in both marine and freshwater species. Infected fish suffer from encephalitis, which leads to abnormal swimming behavior and extensive cellular vacuolation and neuronal degeneration in the central nervous system (CNS) and retina. The marine fish gilthead seabream (Sparus aurata) does not develop VER but it is an asymptomatic carrier of VNNV. In this study, we report that VNNV was able to replicate and persist for up to 3 months in the CNS of the gilthead seabream without causing any neural damage. In addition, we found an early inflammatory response in the CNS that was characterized by the induction of genes encoding pro-inflammatory cytokines, a delayed but persistent induction of anti-inflammatory cytokines, and the infiltration of IgM(+) B lymphocytes, suggesting that local adaptive immunity played a major role in the control of VNNV in the CNS of this species.Developmental and comparative immunology 03/2012; 37(3-4):429-37. · 3.29 Impact Factor -
Article: 17α-Ethynylestradiol alters the immune response of the teleost gilthead seabream (Sparus aurata L.) both in vivo and in vitro.
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ABSTRACT: There is increasing public attention concerning the effect of endocrine disruptor chemicals (EDCs) on the immune system. One important group belonging to EDCs are the environmental estrogens. Commonly found in the effluents in wastewater treatment plants, 17α-ethynylestradiol (EE(2)) which is used in contraceptive pills, is an endocrine disruptor with strong estrogenic effects. This study aims to investigate the capacity of EE(2) to modulate in vivo and in vitro the innate immune response of the gilthead seabream (Sparus aurata L.), a teleost species of great commercial value. For this purpose, adult specimens were bath-exposed to EE(2) (0, 5 and 50 ng/L) and then immunized with hemocyanin in the presence of the adjuvant aluminum. The results indicate that, after 15 days of EE(2)-exposure, the disruptor was able to inhibit in a dose-dependent manner the induction of interleukin-1β (IL-1β) gene expression, but did not significantly alter the specific antibody titer. To shed light on the role played by EE(2) into seabream immune response, leukocytes were exposed in vitro to several concentrations of EE(2) (0, 0.5, 5, 50 and 500 ng/ml) for 3, 16 and 48 h and the production of reactive oxygen intermediates, the phagocytic activity and the gene expression profile of these cells were analyzed. EE(2) was seen to inhibit both cellular activities and to alter the immune gene expression profile in primary macrophages. Thus, low concentrations of EE(2) increase the mRNA levels of IL-1 β, IL-6, tumour necrosis factor α and tumour growth factor β in non-activated macrophages. In contrast, EE(2) treatment of activated macrophages resulted in the decreased expression of pro-inflammatory genes and the increased expression of genes encoding anti-inflammatory and tissue remodeling/repair enzymes. Taken together, our results suggest that EE(2) might alter the capacity of fish to appropriately respond to infection although it does not behave as an immunosuppressor.Developmental and comparative immunology 03/2012; 36(3):547-56. · 3.29 Impact Factor -
Article: Risks of using antifouling biocides in aquaculture.
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ABSTRACT: Biocides are chemical substances that can deter or kill the microorganisms responsible for biofouling. The rapid expansion of the aquaculture industry is having a significant impact on the marine ecosystems. As the industry expands, it requires the use of more drugs, disinfectants and antifoulant compounds (biocides) to eliminate the microorganisms in the aquaculture facilities. The use of biocides in the aquatic environment, however, has proved to be harmful as it has toxic effects on the marine environment. Organic booster biocides were recently introduced as alternatives to the organotin compounds found in antifouling products after restrictions were imposed on the use of tributyltin (TBT). The replacement products are generally based on copper metal oxides and organic biocides. The biocides that are most commonly used in antifouling paints include chlorothalonil, dichlofluanid, DCOIT (4,5-dichloro-2-n-octyl-4-isothiazolin-3-one, Sea-nine 211(®)), Diuron, Irgarol 1051, TCMS pyridine (2,3,3,6-tetrachloro-4-methylsulfonyl pyridine), zinc pyrithione and Zineb. There are two types of risks associated with the use of biocides in aquaculture: (i) predators and humans may ingest the fish and shellfish that have accumulated in these contaminants and (ii) the development of antibiotic resistance in bacteria. This paper provides an overview of the effects of antifouling (AF) biocides on aquatic organisms. It also provides some insights into the effects and risks of these compounds on non-target organisms.International Journal of Molecular Sciences 01/2012; 13(2):1541-60. · 2.60 Impact Factor -
Article: Dietary intake of 17α-ethinylestradiol promotes leukocytes infiltration in the gonad of the hermaphrodite gilthead seabream.
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ABSTRACT: A wide variety of chemicals discharged from industrial and municipal sources have been reported to disrupt the endocrine system of animals, which may be exposed via the food chain and contaminated water. 17α-Ethinylestradiol (EE(2)), a drug used in oral contraceptives and hormone replacement therapy, has a widespread presence in the aquatic environment. Current knowledge on the sensitivity of marine fish to estrogenic environmental chemicals is limited. We report here the effects of dietary intake of EE(2) on gilthead seabream, a marine hermaphrodite teleost, focusing on the immune events that take place in the gonad. When seabream males were fed with 5, 50, 125 and 200μg EE(2)/g food for 7, 14, 21 and 28days an infiltration of acidophilic granulocytes and B lymphocytes occurred in the testis as the same time that spermatogenesis is disrupted. Moreover, the dietary intake of EE(2) promoted a dose-dependent up-regulation of the expression of genes coding for cytokines, chemokines and adhesion molecules correlated with a leukocyte infiltration.Molecular Immunology 08/2011; 48(15-16):2079-86. · 2.90 Impact Factor -
Article: Debaryomyces hansenii L2‐enriched diet enhances the immunity status, gene expression and intestine functionality in gilthead seabream (Sparus aurata L.)
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ABSTRACT: This study evaluates the effects of the dietary administration of the live yeast Debaryomyces hansenii strain L2 on the immune responses of gilthead seabream for 4 weeks. Cellular immune parameters were measured from serum and head-kidney leucocytes respectively. The expression levels of immune-associated genes were quantified by real-time polymerase chain reaction. In addition, the profile of intestinal microbiota was studied by denaturing gradient gel electrophoresis. The results showed that seabream fed a diet containing D. hansenii had significantly increased cellular immune parameters. The yeast-supplemented diet up-regulated the expression of most seabream genes at week 2 and down-regulated all of them at week 4, except in the head-kidney. Finally, a reduction in the diversity of the intestinal microbiota was detected in those specimens receiving the yeast-supplemented diet. These results support the idea that the live yeast D. hansenii strain L2 stimulates the immune system of gilthead seabream.Aquaculture Research 07/2011; · 1.20 Impact Factor -
Article: Molecular and functional characterization of the gilthead seabream β-defensin demonstrate its chemotactic and antimicrobial activity.
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ABSTRACT: Antimicrobial peptides (AMPs) are important mediators of the innate immune response against bacteria and viruses. We have found a β-defensin (BD) gene searching the expressed sequence tags (ESTs) of the teleost fish gilthead seabream (Sparus aurata). The clone contains an open reading frame of 201 bp mRNA that encodes a putative seabream β-defensin (saBD) propeptide of 66 amino acids containing the six conserved cysteines as the main signature of this AMP. The phylogenetic tree shows that saBD, and its fish orthologues, are closely related to the human BD-4. Transcripts of the saBD gene were mainly detected by real-time PCR in the skin, peritoneal leucocytes and head-kidney but scarcely expressed in the peripheral blood. Interestingly, head-kidney leucocytes incubation with synthetic unmethylated CpG oligodeoxynucleotides and bacterial DNA up-regulated the saBD gene expression. Recombinant protein (saBD-V5-His) was expressed in the HEK293 cell line and its functional activity determined. First, seabream head-kidney leucocytes showed chemotactic activity towards supernatants containing saBD-V5-His whilst failed to do so to human recombinant BD-1 y BD-4. Moreover, both cell lysates and supernatants containing saBD-V5-His showed strong antimicrobial activity against Vibrio anguillarum (a seabream pathogenic bacterium) and Bacillus subtilis whilst little on other fish pathogens such as Vibrio harvey and Photobacterium damselae. Further studies will elucidate the existence of other BD genes and their implications on the seabream defense against bacteria and virus.Molecular Immunology 07/2011; 48(12-13):1432-8. · 2.90 Impact Factor
Top co-authors
Top Journals
Institutions
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2001–2013
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Universidad de Murcia
- Facultad de Biología
Murcia, Murcia, Spain
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2012
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Instituto Español de Oceanografia
Madrid, Madrid, Spain
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2009
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Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria
- Centro de Investigación en Sanidad Animal (CISA)
Madrid, Madrid, Spain -
Centro de Investigaciones Biológicas del Noroeste
La Paz, Baja California Sur, Mexico
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2006
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Universidad de Málaga
- Departamento de Microbiología
Málaga, Andalusia, Spain
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