Spatial and temporal variations and assemblage structure of fish species in Beymelek Lagoon,Turkey

Spatial and temporal variations and assemblage structure of fish species were investigated in Beymelek Lagoon, on the south‐western Mediterranean coast of Turkey. A total of 3956 fish, mainly juveniles, from 24 species representing 11 families were sampled by gillnets and trammel nets from February 2006 to January 2007. Twelve of the species were marine straddlers, 11 were marine‐estuarine‐dependent and one was catadromous. Numerical contribution of marine‐estuarine‐dependent species to the total fish abundance was 86.2%, while marine straddlers contributed only 13.8%. Despite the higher number of species, the numerical abundance of marine straddlers was lower than the abundance of marine‐estuarine‐dependent species. The contribution of catadromous species by number was only 0.02%. The assemblage was taxonomically dominated by Sparidae (seven species, 51.3%), Mugilidae (five species, 36.0%), and Clupeidae (one species, 10.3%). Among sparids, Sparus aurata, Diplodus sargus and Lithognathus mormyrus contributed 38.0, 7.2 and 4.6% to the total catch, respectively. The most abundant mugilid species was Liza saliens with 25.2%, followed by Chelon labrosus 5.2%, and Liza aurata 4.0%. Contribution by the other mugilid species to total catch was quite low. The Clupeidae (10.3%) were represented by Sardinella maderensis with 9.1% of the total catch. While the upper reaches of Beymelek Lagoon were dominated by Sparus aurata and Liza saliens, dominant in the lower reaches were Sardinella maderensis, Sparus aurata, Diplodus sargus, Lithognathus mormyrus and Liza saliens. Sparids were generally caught from mid‐summer to mid‐winter while mugilids were caught throughout the year. Clupeids occurred mainly from autumn to spring.     

Development of the first standardised panel of two new microsatellite multiplex PCRs for gilthead seabream (Sparus aurata L.)

The high number of multiplex PCRs developed for gilthead seabream (Sparus aurata L.) from many different microsatellite markers does not allow comparison among populations. This highlights the need for developing a reproducible panel of markers, which can be used with safety and reliability by all users. In this study, the first standardised panel of two new microsatellite multiplex PCRs was developed for this species. Primers of 138 specific microsatellites from the genetic linkage map were redesigned and evaluated according to their genetic variability, allele size range and genotyping reliability. A protocol to identify and classify genotyping errors or potential errors was proposed to assess the reliability of each marker. Two new multiplex PCRs from the best assessed markers were designed with 11 markers in each, named SMsa1 and SMsa2 (SuperMultiplex Sparus aurata). Three broodstocks (59, 47 and 98 breeders) from different Spanish companies, and a sample of 80 offspring from each one, were analysed to validate the usefulness of these multiplexes in the parental assignation. It was possible to assign each offspring to a single parent pair (100% success) using the exclusion method with SMsa1 and/or SMsa2. In each genotyped a reference sample (Ref‐sa) was used, and its DNA is available on request similar to the kits of bin set to genotype by genemapper (v.3.7) software (kit‐SMsa1 and kit‐SMsa2). This will be a robust and effective tool for pedigree analysis or characterisation of populations and will be proposed as an international panel for this species.     

Quantitative trait loci for body growth and sex determination in the hermaphrodite teleost fish Sparus aurata L.

Gilthead sea bream (Sparus aurata L.) is an important marine fish in Mediterranean aquaculture. Sex determination by age and/or body weight is a critical life‐history trait, the genetic basis for which is largely unknown in this sequential hermaphrodite species. Herein, we performed a partial genome scan to map quantitative trait loci (QTL) affecting body weight and sex using 74 informative microsatellite markers from 10 paternal half‐sib families to construct nine linkage groups (LG). In total, four growth‐related QTL (two chromosome‐wide and two genome‐wide) and six QTL related to sex determination (three pairs in three different LGs) were detected (two chromosome‐wide and one genome‐wide). The proportion of phenotypic variation explained by the body‐weight QTL ranged from 9.3% to 17.2%, showing their potential for use in marker‐assisted selection. The results obtained offer solid ground to investigate the structure and function of the genomic regions involved in the mechanisms of sex reversal.     

Additional microsatellites for Sparus aurata and cross‐species amplification within the Sparidae family

Six new microsatellite loci were isolated and characterized in 32 individuals from a farm population of gilthead seabream (Sparus aurata). Expected heterozygosity at all loci was high, ranging from 0.835 to 0.958 with between 10 and 27 alleles per locus. A multiplex polymerase chain reaction protocol was developed using four of the loci. Cross‐species amplification of the loci was tested in six species of the Sparidae family and four loci were successfully amplified in two or more related species.     

Effect of exposure to polycyclic aromatic hydrocarbons on Na +–K +‐ATPase in juvenile gilthead seabream Sparus aurata

In this study, the effects of exposure to pyrene, fluorene and a mixture of selected polycyclic aromatic hydrocarbons (PAH) upon branchial Na ⁺/K ⁺‐ATPase was investigated in juvenile seabream Sparus aurata in a 96 h in vivo experiment. No significant effect was found, and thus it was concluded that branchial Na⁺/K ⁺‐ATPase is probably not affected by PAH.     

The Opecoelidae (Digenea) of sparid fishes of the western Mediterranean. V.Allopodocotyle Pritchard, 1966

Allopodocotyle pedicellata (Stossich, 1887) is redescribed fromSparus aurata from off Corsica and off Provence, France andA. jaffensis (Fischthal, 1980) is redescribed fromSparus pagrus, Dentex dentex andPagellus erythrinus from off Corsica. The status of the genus and these two species is briefly discussed.

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Résumé Allopodocotyle pedicellata (stossich, 1887) est redécrit chezSparus aurata des côtes de la Corse et de la Méditerranée française.A. jaffensis (Fischthal, 1980) est redécrit à partir d'exemplaires provenant deSparus pagrus, Dentex dentex et Pagellus erythrinus des côtes de la Corse. Le statut du genre et de ces deux espèces est discuté.

     

Novel methylation at GpC dinucleotide in the fish Sparus aurata genome

To date, vertebrate DNA has been found methylated at the 5 position of cytosine exclusively in dinucleotide CpG or CpNpG stretches. On the the other hand, we determined that cytosine was methylated unusually in dinucleotide GpC at 5-GGCC-3 sequences in the teleost Sparus aurata EcoRI satellite DNA family. This finding is the first example of methylated GpC sequences in the eukaryotic genomes. At this regard, we have examined the relative methylation levels at this site of the highly repetitive EcoRI satellite DNA family from Sparus aurata different tissues. The EcoRI repeat was remarkably more methylated in male germ cells but hypomethylated in female germ cells at the Hae III restriction site ( GpC). The novel modification and the differential methylation pattern suggest that EcoRI satellite could have a structural and/or functional role at the centromeres of Sparus aurata.     

Efficacy and toxicity of orally administered anticoccidial drugs for innovative treatments of Polysporoplasma sparis (Sitja-Bobadilla and Alvarez-Pellitero 1985) infection in Sparus aurata L.

The purpose of this study was to test experimentally different drugs and therapeutic schemes in order to find a efficient commercial treatment for fish infected with myxosporeans. Two series of land‐based experiments and one experimental cage trial were performed for this purpose. In the first land‐based experiment, 10 and 30 g Sparus aurata naturally infected in the kidneys with Polysporoplasma sparis were used. Initially, six different doses of Fumagillin, two doses of Toltrazuril, and one dose of Amprolium, ESB₃ and Salinomycin were tested. In the second land‐based experiment, 25 and 50 g fish infected with the same parasite were treated with Origanum essential oils, Toltrazuril with propylene glycol, Amprolium, and a combination of Salinomycin 12% + Amprolium (SA). In the field trials, 15 and 155 g S. aurata infected with the same parasite were treated with SA, Origanum essential oils and Fumagillin. In all trials the drugs were incorporated in food and administered according to the selected schemes, while their efficacy was evaluated in terms of mortality (acceptable level <2%), pathology and prevalence rate of P. sparis. According to our results the SA combination proved to be the most effective treatment against P. sparis infection in S. aurata: (i) the therapeutic scheme and commercial product used was not toxic and (ii) a significant reduction in percentage of prevalence was observed.     

Quantitative trait loci for a neurocranium deformity,lack of operculum,in gilthead seabream (Sparus aurata L.)

Lack of operculum, a neurocranial deformity, is the most common external abnormality to be found among industrially produced gilthead seabream (Sparus aurata L.), and this entails significant financial losses. This study conducts, for the first time in this species, a quantitative trait loci (QTL) analysis of the lack of operculum. A total of 142 individuals from a paternal half‐sibling family (six full‐sibling families) were selected for QTL mapping. They had previously shown a highly significant association with the prevalence of lack of operculum in a segregation analysis. All the fish were genotyped for 106 microsatellite markers using a set of multiplex PCRs (ReMsa1–ReMsa13). A linear regression methodology was used for the QTL analysis. Four QTL were detected for this deformity, two of which (QTLOP1 and QTLOP2) were significant. They were located at LG (linkage group) nine and LG10 respectively. Both QTL showed a large effect (about 27%), and furthermore, the association between lack of operculum and sire allelic segregation observed was statistically significant in the QTLOP1 analysis. These results represent a significant step towards including marker‐assisted selection for this deformity in genetic breeding programmes to reduce the incidence of the deformity in the species.     

Localization of corticotropin-releasing factor immunoreactivity in the brain of the teleost Sparus aurata

The distribution of perikarya and fibers containing corticotropin-releasing factor (CRF) was studied in the brain of the teleost Sparus aurata by immunocytochemistry using the peroxidase-antiperoxidase method. Antisera against rat CRF, arginine vasotocin, and human adrenocorticotropin (ACTH) were used. Most CRF-immunoreactive neurons were located in the nucleus lateralis tuberis, but they were absent from the nucleus preopticus, which only contained arginine vasotocin neurons. Few CRF perikarya were identified in the nucleus preopticus periventricularis and in the mesencephalic tegmentum. A conspicuous bundle of immunoreactive fibers ran along the diencephalic floor and pituitary stalk to end near the cells of the hypophysial pars intermedia. No CRF was seen near the adenohypophysial rostral pars distalis. Our results suggest that, in Sparus aurata, CRF is a releasing factor for melanotropic cells. Its role as a releasing factor for ACTH is discussed.     

Pharmacokinetics of cephalexin in sea bream, Sparus aurata (L.), after a single intraperitoneal injection

The pharmacokinetic properties of cephalexin were studied in sea bream (mean weight 77 g), Sparus aurata (L.) after a single intraperitoneal injection (200 mg kg^?1). Pharmacokinetic analysis of the serum concentrations vs time points obtained was performed using non‐compartmental analysis and a compartmental pharmacokinetic model approach. In the latter case, a two‐compartment open model with a lag time gave the best fitting. The maximum peak serum concentration was 5.018 mg ml^?1 kg^?1 1 h post‐treatment. The area under the curve (AUC) cephalexin was 17.394 mg·h kg^?1 and the elimination half‐life of 1.83 h.     

Genetic structure of gilthead seabream, Sparus aurata, in the Central Mediterranean Sea

The gilthead seabream, Sparus aurata, represents an important economic resource for Mediterranean aquaculture. In spite of its wide geographic distribution and economic importance, only recently studies have been carried out on the genetic composition of natural populations, which have revealed a picture of a heterogeneous degree of genetic differentiation among S. aurata populations. In this study an allozyme analysis of samples from six different collecting sites along the Italian and Croatian coasts was carried out, covering an area in the Central Mediterranean sea that has yet to be investigated through gene-enzyme systems. Data on 26 gene loci, 10 of which are polymorphic, indicate a slight but significant genetic structure (F_ST = 0.0167) of the species. A hierarchical analysis of population subdivision made it possible to identify three different assemblages found in the Adriatic Sea, Tyrrhenian Sea and Sardinian Channel, though an isolation by distance model can be rejected. The results are discussed in the light of previous literature and taking conservation into consideration.     

The genetic population structure and temporal genetic stability of gilthead sea bream Sparus aurata populations in the Aegean and Ionian Seas,using microsatellite DNA markers

We examined 662 gilthead sea bream Sparus aurata from wild samples of the species in the Aegean and Ionian Seas, using 20 EST-linked microsatellite markers, in three multiplex panels, as well as seven anonymous loci. Most of the markers were revealed to be highly polymorphic. We found low genetic differentiation between the sampling stations/areas with total F_ST 0.002 (P < 0.05). Based on comparison of five temporal samples, our results indicate genetic data consistency over time for all tested samples, pointing to stable populations, despite reported repeated escape events. Our results confirm the genetic population structure previously observed in these specific areas, using by far more markers than in previous studies in both coding and non-coding DNA loci. The limited genetic structure and the temporal genetic stability indicate neither major genetic differentiation of local populations by geographic isolation nor influence from anthropogenic factors. These results provide a baseline for future reference in any management programme of both wild and farmed population of S. aurata as well as of other aquaculture species with a potential introgression among farmed and wild populations.     

Retinal development in the gilthead seabream Sparus aurata

The retinal development of the gilthead seabream Sparus aurata has been analysed from late embryonic development to juvenile stages using classical histological and immunohistological methods. Five significant phases were established. Phases 1 and 2 comprise the late embryonic and hatching stages, respectively. The results indicate that during these early stages the retina is composed of a single neuroblastic layer that consists of undifferentiated retinal progenitor cells. Phase 3 (late prolarval stage) is characterized by the emergence of the retinal layers and the appearance of neurochemical profiles in differentiating photoreceptors, amacrine and ganglion cells. Phases 4 and 5 comprise the late larval and juvenile stages. In these stages, all the retinal cell types can be detected immunohistochemically. All the maturational events described are first detected in the central retina and, as development progresses, spread to the rest of the retina following a central‐to‐peripheral gradient. The results of this study suggest that S. aurata is an altricial teleost species that hatches with a morphologically undifferentiated retina. The most relevant processes involved in retinogenesis occur during the late prolarval stage (phase 3).     

Length–weight relationships for 22 fish species of the Mar Menor coastal lagoon (western Mediterranean Sea)

Length–weight relationships were estimated for 22 species of the Mar Menor coastal lagoon (south‐eastern Spain): Lipophrys dalmatinus, Salaria pavo, Callionymus pusillus, Aphanius iberus, Engraulis encrasicolus, Gobius cobitis, G. niger, G. paganellus, Pomatoschistus marmoratus, Symphodus cinereus, Chelon labrosus, Liza aurata, L. ramado, L. saliens, Mugil cephalus, Diplodus puntazzo, D. sargus sargus, Lithognathus mormyrus, Sarpa salpa, Sparus aurata, Hippocampus guttulatus and Syngnathus abaster. The captures were made in shallow areas of the lagoon during 2002–2004. Significant length–weight relationships were found for all species.     

A Microsatellite Linkage Map of Striped Bass (Morone saxatilis) Reveals Conserved Synteny with the Three-Spined Stickleback (Gasterosteus aculeatus)

The striped bass (Morone saxatilis) and its relatives (genus Morone) are of great importance to fisheries and aquaculture in North America. As part of a collaborative effort to employ molecular genetics technologies in striped bass breeding programs, we previously developed nearly 500 microsatellite markers. The objectives of this study were to construct a microsatellite linkage map of striped bass and to examine conserved synteny between striped bass and three-spined stickleback (Gasterosteus aculeatus). Of 480 microsatellite markers screened for polymorphism, 289 informative markers were identified and used to genotype two half-sib mapping families. Twenty-six linkage groups were assembled, and only two markers remain unlinked. The sex-averaged map spans 1,623.8 cM with an average marker density of 5.78 cM per marker. Among 287 striped bass microsatellite markers assigned to linkage groups, 169 (58.9%) showed homology to sequences on stickleback chromosomes or scaffolds. Comparison between the stickleback genome and the striped bass linkage map revealed conserved synteny between these two species. This is the first linkage map for any of the Morone species. This map will be useful for molecular mapping and marker-assisted selection of genes of interest in striped bass breeding programs. The conserved synteny between striped bass and stickleback will facilitate fine mapping of genome regions of interest and will serve as a new resource for comparative mapping with other Perciform fishes such as European sea bass (Dicentrarchus labrax), gilthead sea bream (Sparus aurata), and tilapia (Oreochromis ssp.).     

On the first record of a potentially harmful fish, Sparus aurata in the Gulf of California

The record of free and healthy Mediterranean gilt-head seabream Sparus aurata Linnaeus, 1758 in waters of the Gulf of California, México is documented for the first time. Consequences of this introduction are briefly outlined.     

In vitro effect of cortisol and urotensin I on arginine vasotocin and isotocin secretion from pituitary cells of gilthead sea bream Sparus aurata

This study aimed at determining whether in vitro secretion of two neuropeptides, arginine vasotocin (AVT) and isotocin (IT), from pituitary cells of gilthead sea bream Sparus aurata was affected by cortisol and urotensin (UI). Pituitary cells were exposed to 1·4 × 10^?8, 1·4 × 10^?7 and 0·4 × 10^?6 M cortisol and 10^?12, 10^?10 and 10^?8 M UI for 6, 24 and 48 h, respectively. AVT and IT contents were determined in the culture media by high‐performance liquid chromatography (HPLC). An increase in AVT secretion and a decrease in IT secretion were observed at all cortisol doses. UI increased AVT secretion after 6 h of incubation at all doses. After 24 h, however, only the highest dose of UI still displayed an effect. IT secretion was not influenced by UI. It was thus demonstrated that cortisol does influence AVT and IT secretion from S. aurata pituitary cells, while UI regulates AVT secretion, as a component of hypothalamic–pituitary–interrenal (HPI) axis in this species.     

Influence of environmental conditions on demand-feeding behaviour of gilthead seabream (Sparus aurata)

We studied the demand‐feeding behaviour was studied of gilthead seabream (Sparus aurata, L.) reared under either constant (25 ± 0.5°C, 12 : 12 L : D, control group) or natural (experimental group) temperature and photoperiod conditions during a period from winter to summer. Hourly demand‐feeding activity profiles were recorded using self‐feeding devices; these profiles showed that control group behaved entirely as a diurnal species, exhibiting no nightly activity and decreased demand rates in winter months. The experimental group did exhibit nightly activity (in incomplete darkness); this group also showed reduced demand rates in winter months, accompanied by a demand peak shift towards evening/night hours that followed the day's temperature peak of colder months.     

Visualizing mineralization in deformed opercular bones of larval gilthead sea bream (Sparus aurata)

During the rearing process of gilthead sea bream (Sparus aurata), abnormal development of the opercular bone is particularly common (Aquaculture 156, 1997, 165). In order to alleviate its occurrence in rearing facilities, it’s crucial to identify the very first physical signs of deviation in normal skeletal development. Nano‐CT‐scanning was tested for its applicability to quantify deviations in bone mineralization levels. Seven opercles were dissected from larvi of 65 days post hatching, randomly sampled at the commercial sea bream hatchery Maricoltura di Rosignano Solvay (Livorno, Italy). The samples were nano‐CT‐scanned and computationally reconstructed. Mineralization intensity was colorcoded using Amira software, resulting in a detailed visualization of opercular morphology and mineralization patterns. In conclusion, nano‐CT‐scanning promises to be a good tool to both describe morphology and detect mineralization levels in the early onset of deformities.