IFAPA Centro El Toruno

El Puerto de Santa María, Spain

IFAPA Centro El Toruno

El Puerto de Santa María, Spain
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Souto S.,University of Santiago de Compostela | Lopez-Jimena B.,IFAPA Centro El Toruno | Lopez-Jimena B.,University of Stirling | Alonso M.C.,University of Malaga | And 2 more authors.
Veterinary Microbiology | Year: 2015

The susceptibility of juvenile European sea bass and Senegalese sole to three VNNV isolates (a reassortant RGNNV/SJNNV, as well as the parental RGNNV and SJNNV genotypes) has been evaluated by challenges using two inoculation ways (bath and intramuscular injection). The results demonstrate that these two fish species are susceptible to all the VNNV isolates tested. In European sea bass, RGNNV caused the highest cumulative mortality, reaching maximum values of viral RNA and titres. Although the SJNNV isolate did not provoke mortality or clinical signs of disease in this fish species, viral production in survivor fish was determined; on the other hand the reassortant isolate did cause mortality and clinical signs of disease, although less evident than those recorded after RGNNV infection. These results suggest that the changes suffered by the SJNNV RNA2 segment of the reassortant isolate, compared to the parental SJNNV, may have involved host-specificity and/or virulence determinants for European sea bass. Regarding Senegalese sole, although the three isolates caused 100% mortality, the reassortant strain provoked the most acute symptoms, and more quickly, especially in the bath challenge. This was also the isolate showing less difference between the number of RNA copies and viral titre, reaching the highest titres of infective viral particles in nervous tissue of infected animals. The RGNNV isolate produced the lowest values of infective viral particles. All these results suggest that the RGNNV and the reassortant isolates are the most suited for infecting European sea bass and Senegalese sole, respectively. © 2015 Elsevier B.V.

Fernandez-Diaz C.,IFAPA Centro El Toruno | Coste O.,IFAPA Centro El Toruno | Malta E.-J.,IFAPA Centro Agua del Pino
Algal Research | Year: 2017

The potential of chitosan-based nanoparticles to incorporate ulvan was investigated and the immunostimulant properties of ulvan polysaccharide as nanocarrier in Solea senegalensis macrophage were evaluated. Ulvan native extract (UL) and ulvan fractionated by chemical (ULQ) or enzymatic (ULE) methods were obtained from Ulva ohnoi macroalgae in culture. These different ulvan types were tested for their ability to form polymeric nanoparticles. Nanoparticles including either unaltered or fractionated ulvan were prepared by ionotropic gelation and characterized. Activity of the ulvan-loaded nanoparticles was tested in vitro on fish macrophages against free unaltered or fractioned ulvan extract. Intracellular and extracellular reactive oxygen species (ROS) production from the different ulvan products was determined in S. senegalensis macrophages using oxidative burst assay. The native ulvan extract (UL) as well as the fractioned form (ULQ and ULE) successfully yielded nanoparticles with a size of 250–300 nm and with a Z-potential of 30–40 mV. Highest nanoparticle production was achieved using ulvan native extract (NPsUL). Our results indicate that the configuration of the structure of ulvan molecules influence the immune response; in this way, unaltered ulvan is required for successful stimulation of Solea macrophages by Ulva ohnoi polysaccharides. This immune response significantly increases when unaltered ulvan is nanoencapsulated. We conclude that the here developed hybrid polysaccharide nanoparticles composed of chitosan and ulvan are functional. This might open the way for production of nanocarriers that can be used for oral administration of active compounds in aquaculture. © 2017

Blanco-Vives B.,University of Murcia | Aliaga-Guerrero M.,University of Cádiz | Canavate J.P.,IFAPA Centro El Toruno | Munoz-Cueto J.A.,University of Cádiz | Sanchez-Vazquez F.J.,University of Murcia
Chronobiology International | Year: 2011

Light plays a key role in the development of biological rhythms in fish. Previous research on Senegal sole has revealed that both spawning rhythms and larval development are strongly influenced by lighting conditions. However, hatching rhythms and the effect of light during incubation are as yet unexplored. Therefore, the aim of this study was to investigate the impact of the light spectrum and photoperiod on Solea senegalensis eggs and larvae until day 7 post hatching (dph). To this end, eggs were collected immediately after spawning during the night and exposed to continuous light (LL), continuous darkness (DD), or light-dark (LD) 12L:12D cycles of white light (LDW), blue light (LDB; λpeak = 463 nm), or red light (LDR; λpeak = 685 nm). Eggs exposed to LDB had the highest hatching rate (94.5% ± 1.9%), whereas LDR and DD showed the lowest hatching rate (54.4% ± 3.9% and 48.4% ± 4.2%, respectively). Under LD conditions, the hatching rhythm peaked by the end of the dark phase, but was advanced in LDB (zeitgeber time 8 [ZT8]; ZT0 representing the onset of darkness) in relation to LDW and LDR (ZT11). Under DD conditions, the same rhythm persisted, although with lower amplitude, whereas under LL the hatching rhythm split into two peaks (ZT8 and ZT13). From dph 4 onwards, larvae under LDB showed the best growth and quickest development (advanced eye pigmentation, mouth opening, and pectoral fins), whereas larvae under LDR and DD had the poorest performance. These results reveal that developmental rhythms at the egg stage are tightly controlled by light characteristics, underlining the importance of reproducing their natural underwater photoenvironment (LD cycles of blue wavelengths) during incubation and early larvae development of fish. © Informa Healthcare USA, Inc.

Marin-Juez R.,University of Barcelona | Castellana B.,University of Barcelona | Manchado M.,IFAPA Centro El Toruno | Planas J.V.,University of Barcelona
General and Comparative Endocrinology | Year: 2011

In male teleosts, testicular steroids are essential hormones for the regulation of spermatogenesis and their production is regulated by pituitary gonadotropins. In the Senegalese sole (Solea senegalensis), an economically important flatfish with semi-cystic and asynchronous spermatogenesis, the gonadotropic regulation of spermatogenesis, particularly regarding the production and regulation of testicular steroids, are not well understood. For this reason, we first cloned and characterized the response of several key genes for the production and action of testicular steroids to the in vivo administration of human chorionic gonadotropin (hCG) and, second, we investigated the transcriptomic effects of hCG in the Senegalese sole testis. We succeded in cloning the full-length cDNAs for Steroidogenic Acute Regulatory protein (StAR), 3β-hydroxysteroid dehydrogenase (3β-HSD), 17β-HSD and 20β-HSD and a partial cDNA for the nuclear progesterone receptor. In this study we also identified a transcript encoding a protein with homology to StAR, which we named StAR-like, that could represent a new member of the StAR-related lipid transfer (START) family. All the cloned genes were expressed in the testis and their expression levels were significantly increased by the in vivo administration of hCG. The plasma levels of testosterone and 11-ketotestosterone also increased in response to hCG administration, likely as a result of the induction of the expression of steroidogenic enzymes by hCG. Furthermore, gene expression analysis by microarray identified 90 differentially expressed genes in the testis in response to hCG administration, including genes potentially involved in steroidogenesis, progression of spermatogenesis and germ cell maturation and cytoskeletal organization. Our results have identified for the first time a number of key genes involved in the regulation of steroid production and spermatogenesis in the Senegalese sole testis that are under gonadotropic control. © 2011 Elsevier Inc.

Cerda J.,CSIC - Institute of Marine Sciences | Manchado M.,IFAPA Centro El Toruno
Genes and Nutrition | Year: 2013

Fish aquaculture is considered to be one of the most sustainable sources of protein for humans. Many different species are cultured worldwide, but among them, marine flatfishes comprise a group of teleosts of high commercial interest because of their highly prized white flesh. However, the aquaculture of these fishes is seriously hampered by the scarce knowledge on their biology. In recent years, various experimental 'omics' approaches have been applied to farmed flatfishes to increment the genomic resources available. These tools are beginning to identify genetic markers associated with traits of commercial interest, and to unravel the molecular basis of different physiological processes. This article summarizes recent advances in flatfish genomics research in Europe. We focus on the new generation sequencing technologies, which can produce a massive amount of DNA sequencing data, and discuss their potentials and applications for de novo genome sequencing and transcriptome analysis. The relevance of these methods in nutrigenomics and foodomics approaches for the production of healthy animals, as well as high quality and safety products for the consumer, is also briefly discussed. © 2012 Springer-Verlag.

Vinas J.,CSIC - Institute of Marine Sciences | Vinas J.,University of Girona | Asensio E.,IFAPA Centro El Toruno | Canavate J.P.,IFAPA Centro El Toruno | Piferrer F.,CSIC - Institute of Marine Sciences
Aquaculture | Year: 2013

Like other flatfishes, the Senegalese sole exhibits sexual growth dimorphism in favor of females and thus all-female stocks would be desirable. However, reproduction-associated problems persist and basic aspects of its biology such as sex determination and differentiation are still unknown. In this study, we histologically examined the process of gonadal sex differentiation in the Senegalese sole and tested the effects of treatment with sex steroids and an aromatase inhibitor on sex ratios. Sex differentiation in females started before 98 days post fertilization (dpf), when fish had an average total length (TL) of ~ 33 mm. In contrast, males began sex differentiation by 127 dpf with a mean TL of ~ 44 mm. At this time, an incipient growth advantage in favor of females was already observed. All fish were sexually differentiated by ~ 48 mm TL (~ 170. dpf). Treatment between 98 and 169 dpf with the synthetic androgen 17α-methyldihydrotestosterone (10 mg/kg) or the aromatase inhibitor fadrozole (100. mg/kg) significantly increased the proportion of males up to ~ 95% while the natural estrogen, estradiol-17β (10 mg/kg), increased the proportion of females by about 50% but reduced growth with respect to the control values. Together, these results show that the Senegalese sole is a differentiated gonochoristic species in which gonads start to form within the first three months and sex differentiation is completed after animals reach their fifth month of age, with females differentiating earlier than males. Results also indicate that the labile period starts at < 98. dpf and lasts until ~ 125. dpf. These results show that sex control can be achieved in the Senegalese sole, an important step towards the establishment of all-female stocks to improve the currently compromised production of this economically important species. © 2012 Elsevier B.V.

Ponce M.,IFAPA Centro El Toruno | Infante C.,IFAPA Centro El Toruno | Manchado M.,IFAPA Centro El Toruno
General and Comparative Endocrinology | Year: 2010

Thyroid hormones (THs) play a key role in larval development, growth and metamorphosis in flatfish. Their synthesis is tightly regulated by the hypothalamic-pituitary-thyroid axis. Thyroid-stimulating hormone receptor (TSHR) is a key protein in the control of thyroid function stimulating TH synthesis after binding its ligand, the thyrotropin. In teleost fish, numerous reports have associated the TSHR with gametogenesis. However, little information about its role during larval development is available. In this study, we report the cloning of two different cDNAs with high similarity to TSHR. Phylogenetic analysis clustered both cDNAs separately. One of them (referred to TSHR) grouped with TSHR orthologs in tetrapods and teleost fish and possessed the three typical conserved domains and regulatory motifs. The second receptor (referred to as TSHRtr-like) represented a novel truncated cDNA bearing the extracellular and part of the transmembrane domain. TSHRtr-like orthologs were only found in teleosts, which suggests that it could have appeared after fish-specific 3R genome duplication. Expression profiles of both genes are analyzed in juvenile tissues and during larval development using a real-time PCR approach. In juvenile fish, TSHR and TSHRtr-like are expressed ubiquitously although transcript levels varied between organs. In both cases, the highest mRNAs levels are detected in brain. During larval development, both genes are expressed to a high level during the first stages (2-3 days after hatching) reducing progressively their abundance in the whole larvae during metamorphosis. This reduction in mRNA abundance is more accentuated for the TSHRtr-like gene. To evaluate the possible regulation of both receptors by T4 during sole metamorphosis, larvae are exposed to the goitrogen thiourea (TU). Only TSHRtr-like modifies its expression, increasing its transcripts at 11 days after treatment. Moreover, adding exogenous T4 hormone to TU-treated larvae restores the TSHRtr-like steady-state levels similar to the untreated control. Overall, these results demonstrate the existence of two thyrotropin receptors differentially regulated by THs in teleosts. © 2010 Elsevier Inc.

Mackerels of the genus Scomber are commercially important species, but their taxonomic status is still controversial. Although previous phylogenetic data support the recognition of Atlantic Scomber colias and Pacific Scomber japonicus as separate species, it is only based on the analysis of partial mitochondrial and nuclear DNA sequences. In an attempt to shed light on this relevant issue, we have determined the complete mitochondrial DNA sequence of S. colias, S. japonicus, and Scomber australasicus. The total length of the mitogenomes was 16,568 bp for S. colias and 16,570 bp for both S. japonicus and S. australasicus. All mitogenomes had a gene content (13 protein-coding, 2 rRNAs, and 22 tRNAs) and organization similar to that observed in Scomber scombrus and most other vertebrates. The major noncoding region (control region) ranged between 865 and 866 bp in length and showed the typical conserved blocks. Phylogenetic analyses revealed a monophyletic origin of Scomber species with regard to other scombrid fish. The major finding of this study is that S. colias and S. japonicus were significantly grouped in distinct lineages within Scomber cluster, which phylogenetically constitutes evidence that they may be considered as separate species. Additionally, molecular data here presented provide a useful tool for evolutionary as well as population genetic studies. © 2009 Elsevier B.V.

Infante C.,IFAPA Centro El Toruno | Ponce M.,IFAPA Centro El Toruno | Manchado M.,IFAPA Centro El Toruno
Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology | Year: 2011

Calsequestrin is a moderate-affinity, high-capacity Ca 2+ binding protein in the sarcoplasmic reticulum of skeletal and cardiac muscle that seems to act as an intralumenal Ca 2+ buffer. Two different isoforms have been described in mammals, the skeletal and cardiac isoforms, encoded by CASQ1 and CASQ2 genes, respectively. In this study, we present molecular phylogenetic evidence of a gene duplication event of both calsequestrin genes in teleosts, referred to as casq1a/casq1b and casq2a/casq2b. We obtained the entire cDNAs encoding the four genes in the Senegalese sole (Solea senegalensis Kaup). Main features and sequence identities with other fish and mammalian calsequestrins are described. Expression profiles during larval development and in juvenile tissues were analyzed using a real-time PCR approach. In juvenile fish, casq1a and casq1b were highly expressed in skeletal muscle, whereas the highest casq2a and casq2b transcript levels were detected in heart and brain, respectively. During metamorphosis, casq2a and casq1b expression remained unchanged. In contrast, casq1a and casq2b mRNAs exhibited a continuous increase from the beginning of metamorphosis until post-metamorphosis. Transcriptional regulation of casq1 and casq2 genes by thyroid hormones (THs) was also evaluated. Larvae exposed to the goitrogen thiourea (TU) exhibited higher casq1a mRNA levels than untreated control, whereas expression of the remaining genes did not vary significantly. Moreover, addition of exogenous T4 hormone to TU-treated larvae increased the casq1a steady-state levels with respect to the untreated control at metamorphosis climax. Comprehensively, these results demonstrate the existence of four calsequestrin genes in teleosts differentially regulated by THs. © 2011 Elsevier Inc.

Gonzalez-Ortegon E.,Bangor University | Gonzalez-Ortegon E.,IFAPA Centro El Toruno | Gimenez L.,Bangor University
Marine Ecology Progress Series | Year: 2014

We studied the effects of environmental conditions experienced during embryonic and larval phases on development and larval survival of the marine shrimp Palaemon serratus, and examined how these conditions modified the relationship between larval and maternal pheno types. Egg-carrying females were incubated at different temperatures (12 and 18°C), and freshly hatched larvae were exposed to a combination of temperatures (18 and 24°C), salinities (25 and 32 PSU) and food conditions (ad libitum vs. limited). Temperatures experienced by embryos had no significant effects on development, and only weak effects on survival, whereas environmental conditions experienced by larvae had strong effects on development-the duration of development was longer at lower temperatures and under food-limited conditions, and food limitation increased the number of larval instars necessary to reach the juvenile phase (especially at the highest temperature), perhaps reflecting a mismatch between increased metabolic demands and reduced energy supply. Links between larval and female phenotypes were evident: large females generally produced significantly larger larvae than smaller females. In larvae reared under food limitation, average development time and number of instars required to reach the juvenile phase were negatively correlated with average larval body mass at hatching. Thus, larval development is linked to initial larval body mass and female body size; however, these links can be modified by environmental conditions experienced by the larvae. In situations of high temperatures and food limitation, larger P. serratus females may play a more important role in the maintenance of populations, as they produce large offspring capable of ameliorating the effects of temperature and food limitation on development. © Inter-Research 2014.

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