Detection of growth-related QTLs in turbot (Scophtalmus maximux)

Background The turbot (Scophthalmus maximus) is a highly appreciated European aquaculture species. Growth related traits constitute the main goal of the ongoing genetic breeding programs of this species. The recent construction of a consensus linkage map in this species has allowed the selection of a panel of 100 homogeneously distributed markers covering the 26 linkage groups (LG) suitable for QTL search. In this study we addressed the detection of QTL with effect on body weight, length and Fulton's condition factor. Results Eight families from two genetic breeding programs comprising 814 individuals were used to search for growth related QTL using the panel of microsatellites available for QTL screening. Two different approaches, maximum likelihood and regression interval mapping, were used in order to search for QTL. Up to eleven significant QTL were detected with both methods in at least one family: four for weight on LGs 5, 14, 15 and 16; five for length on LGs 5, 6, 12, 14 and 15; and two for Fulton's condition factor on LGs 3 and 16. In these LGs an association analysis was performed to ascertain the microsatellite marker with the highest apparent effect on the trait, in order to test the possibility of using them for marker assisted selection. Conclusions The use of regression interval mapping and maximum likelihood methods for QTL detection provided consistent results in many cases, although the high variation observed for traits mean among families made it difficult to evaluate QTL effects. Finer mapping of detected QTL, looking for tightly linked markers to the causative mutation, and comparative genomics are suggested to deepen in the analysis of QTL in turbot so they can be applied in marker assisted selection programs

Hatchery manual for broodstock management and larval production of tubrot (Psetta maxima)

This hatchery manual is intended to provide detailed information from available published work and grey literature on turbot broodstock management and larval production. In reviewing larviculture techniques for turbot, it is notable that the major initial zoo technical advances were made in the 1980s. Subsequent refinements have been industry-led and are subject to commercial confidentiality. Some actors in the sector that have been approached either did not provide requested information or denied access to their sources of information. This manuscript therefore considers those aspects of commercial rearing techniques that are in the public domain, together with the applied scientific literature and information collected from different experts. With the aim to cover all aspects related to the production of turbot juveniles in Dutch farms from egg to fish of 10-15 g, the manual describes in details different steps. These have been grouped around the broodstock management, the hatchery/nursery period, the on growing and the grow out periods. A list of common diseases in turbot has been added at the end. The manual provides a link to the most update information available on live prey production and enrichment

Non-synonymous variation and protein structure of candidate genes associated with selection in farm and wild populations of turbot (Scophthalmus maximus)

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The influence of anaerobic muscle activity, maturation and season on the flesh quality of farmed turbot

In order to test seasonal, rearing, maturing and anaerobic muscle activity effect on the flesh quality of turbot (Scophthalmus maximus) a total of 80 farmed turbot from three different strains from reared under natural or continuous light were killed by a percussive blow to the head in November (winter, Icelandic strain), March (spring, Portuguese strain) and June (summer, domesticated strain (France turbot)). To test the effect of anaerobic muscle activity, 10 fish were on each occasion pre rigor filleted, where one fillet was used as a control, while the other fillet was electrically stimulated using a squared 5 Hz, 10 V pulsed DC for 3 min. All pre rigor fillets were measured for pH, weighed, wrapped in aluminum foil and stored in polystyrene boxes with ice. After 7 days of storage the fillets were measured instrumentally for pH, drip loss, colour (CIE L* a* b*) and texture properties such as hardness and shear force, while fillet shrinkage and colour (RBG) were evaluated with computer imaging on photographs from a standard lightbox. Results showed that softness of the flesh was mainly influenced by factors associated with growth, such as season, photoperiod and maturation. Anaerobic muscle activity simulated with electrical stimulation caused an increase in drip loss (<1%) and loss of shear force (<4%), but had no effect on hardness or fillet shrinkage. Computer imaging revealed that muscle contractions related to the electrical stimulus forced out blood from the fillet causing less reddishness for the entire storage period. We conclude that a pH drop upon slaughter associated with anaerobic muscle activity has a minor effect on the flesh quality in the short run, while seasonal/alternatively genetic effects are predominant

Integration of host-pathogen functional genomics data into the chromosome-level genome assembly of turbot (Scophthalmus maximus)

Disease resilience is of utmost relevance for turbot aquaculture. Several infective diseases, covering a broad spectrum from viruses, bacteria to different parasites, have been identified by industry. Since they increase mortality rates, reduce feed conversion ratios and slow down growth rate, genetic breeding programs for increasing disease resilience are recognized as a useful alternative for controlling pathologies. For this, knowledge of the genetic basis underlying resilience using genomic tools is essential to develop the best effective breeding strategies. In the present study, we compiled the existing genomic information generated in the last decade to construct an integrated atlas of candidate genes and genomic regions involved in pathogen resistance against the main turbot industrial pathogens (Aeromonas salmonicida, Philasterides dicentrarchi, Enteromyxum scophthalmi and the VHS virus) within the chromosome-level turbot genome assembly recently released. Information comprehends reannotated differentially expressed genes (DEG) in different tissues along temporal series, QTL markers associated with important productive traits (disease resistance and growth) and signatures of domestic or wild selection, represented by runs of homozygosity (ROHi) islands and outlier markers for divergent selection. Most genetic features were successfully relocated in the turbot assembly including 81.1% of the total DEGs, plus all QTL markers, ROHi and outlier markers. The updated annotation of DEGs for resistance to each pathology demonstrated significant changes. While the new annotation of 53–83% of the DEGs was coherent with the original, roughly 10–24% showed imprecise annotations in both assembly versions, ∼5% lost their original annotation and 2–24% were now annotated. Functional enrichment revealed mostly functions related to immune response, such as chemotaxis, apoptosis regulation, leukocyte differentiation, cell adhesion, iron homeostasis and vascular permeability. Some DEGs, such as celsr1a (cadherin EGF LAG seen-pass G-type receptor 1), fgg (fibrinogen gamma chain) and c1qtnf9 (C1q and TNF related 9) were found near pathogen-associated QTL markers. Also, some shared DEGs for resistance to all pathogens were positioned near QTL markers or ROHi, such as hamp (hepcidin-1), plg (plasminogen) and a fibrinogen alpha chain-like gene. Overall, our results provide an integrative insight into the genetic architecture of turbot response to a range of pathogens that could prove useful for future genomic studies to benefit aquaculture breeding programsS

An Expressed Sequence Tag (EST)-enriched genetic map of turbot (Scophthalmus maximus): a useful framework for comparative genomics across model and farmed teleosts

[Background] The turbot (Scophthalmus maximus) is a relevant species in European aquaculture. The small turbot genome provides a source for genomics strategies to use in order to understand the genetic basis of productive traits, particularly those related to sex, growth and pathogen resistance. Genetic maps represent essential genomic screening tools allowing to localize quantitative trait loci (QTL) and to identify candidate genes through comparative mapping. This information is the backbone to develop marker-assisted selection (MAS) programs in aquaculture. Expressed sequenced tag (EST) resources have largely increased in turbot, thus supplying numerous type I markers suitable for extending the previous linkage map, which was mostly based on anonymous loci. The aim of this study was to construct a higher-resolution turbot genetic map using EST-linked markers, which will turn out to be useful for comparative mapping studies. [Results] A consensus gene-enriched genetic map of the turbot was constructed using 463 SNP and microsatellite markers in nine reference families. This map contains 438 markers, 180 EST-linked, clustered at 24 linkage groups. Linkage and comparative genomics evidences suggested additional linkage group fusions toward the consolidation of turbot map according to karyotype information. The linkage map showed a total length of 1402.7 cM with low average intermarker distance (3.7 cM; ~2 Mb). A global 1.6:1 female-to-male recombination frequency (RF) ratio was observed, although largely variable among linkage groups and chromosome regions. Comparative sequence analysis revealed large macrosyntenic patterns against model teleost genomes, significant hits decreasing from stickleback (54%) to zebrafish (20%). Comparative mapping supported particular chromosome rearrangements within Acanthopterygii and aided to assign unallocated markers to specific turbot linkage groups. [Conclusions] The new gene-enriched high-resolution turbot map represents a useful genomic tool for QTL identification, positional cloning strategies, and future genome assembling. This map showed large synteny conservation against model teleost genomes. Comparative genomics and data mining from landmarks will provide straightforward access to candidate genes, which will be the basis for genetic breeding programs and evolutionary studies in this species.This study was supported by the projects: Consolider Ingenio Aquagenomics (CSD200700002), Spanish Ministerio de Ciencia e Innovación (AGL2009-13273), and Xunta de Galicia local Government (09MMA011261PR). We are indebted to Lucía Insua, María Portela, Susana Sánchez, María López, Mónica Otero and Sonia Gómez for technical assistance. B.G. Pardo was supported by an Isidro Parga Pondal research fellowship from Xunta de Galicia (Spain)

Sustainable fish feeds: potential of emerging protein sources in diets for juvenile turbot (Scophthalmus maximus) in RAS

In Europe, turbot aquaculture has a high potential for sustainable production, but the low tolerance to fishmeal replacement in the diet represents a big issue. Therefore, this study investigated the effects of more sustainable feed formulations on growth and feed performance, as well as nutritional status of juvenile turbot in recirculating aquaculture systems. In a 16-week feeding trial with 20 g juvenile turbot, one control diet containing traditional fishmeal, fish oil and soy products and two experimental diets where 20% of the fishmeal was replaced either with processed animal proteins (PAP) or with terrestrial plant proteins (PLANT) were tested. Irrespective of diets, growth performance was similar between groups, whereas the feed performance was significantly reduced in fish of the PAP group compared to the control. Comparing growth, feed utilisation and biochemical parameters, the results indicate that the fish fed on PAP diet had the lowest performance. Fish fed the PLANT diet had similar feed utilisation compared to the control, whereas parameters of the nutritional status, such as condition factor, hepato-somatic index and glycogen content showed reduced levels after 16 weeks. These effects in biochemical parameters are within the physiological range and therefore not the cause of negative performance. Since growth was unaffected, the lower feed performance of fish that were fed the PAP formulation might be balanced by the cost efficient formulation in comparison to the commercial and the PLANT formulations. Present study highlights the suitability of alternative food formulation for farmed fish

Consolidation of the genetic and cytogenetic maps of turbot (Scophthalmus maximus) using FISH with BAC clones

This is a post-peer-review, pre-copyedit version of an article published in Chromosoma. The final authenticated version is available online at: https://doi.org/10.1007/s00412-014-0452-2Bacterial artificial chromosomes (BAC) have been widely used for fluorescence in situ hybridization (FISH) mapping of chromosome landmarks in different organisms, including a few in teleosts. In this study, we used BAC-FISH to consolidate the previous genetic and cytogenetic maps of the turbot (Scophthalmus maximus), a commercially important pleuronectiform. The maps consisted of 24 linkage groups (LGs) but only 22 chromosomes. All turbot LGs were assigned to specific chromosomes using BAC probes obtained from a turbot 5× genomic BAC library. It consisted of 46,080 clones with inserts of at least 100 kb and <5 % empty vectors. These BAC probes contained gene-derived or anonymous markers, most of them linked to quantitative trait loci (QTL) related to productive traits. BAC clones were mapped by FISH to unique marker-specific chromosomal positions, which showed a notable concordance with previous genetic mapping data. The two metacentric pairs were cytogenetically assigned to LG2 and LG16, and the nucleolar organizer region (NOR)-bearing pair was assigned to LG15. Double-color FISH assays enabled the consolidation of the turbot genetic map into 22 linkage groups by merging LG8 with LG18 and LG21 with LG24. In this work, a first-generation probe panel of BAC clones anchored to the turbot linkage and cytogenetical map was developed. It is a useful tool for chromosome traceability in turbot, but also relevant in the context of pleuronectiform karyotypes, which often show small hardly identifiable chromosomes. This panel will also be valuable for further integrative genomics of turbot within Pleuronectiformes and teleosts, especially for fine QTL mapping for aquaculture traits, comparative genomics, and whole-genome assemblyThis study was supported by Spain’s Ministerio de Ciencia e Innovación (AGL2009-13273), Consolider Ingenio Aquagenomics (CSD200700002) and Xunta de Galicia (09MMA011261PR; 10MMA200027PR). Samples for cytogenetic analysis were kindly supplied by Cluster de Acuicultura de GaliciaS

Development and Validation of Single Nucleotide Polymorphisms (SNPs) Markers from Two Transcriptome 454-Runs of Turbot (Scophthalmus maximus) Using High-Throughput Genotyping

The turbot (Scophthalmus maximus) is a commercially valuable flatfish and one of the most promising aquaculture species in Europe. Two transcriptome 454-pyrosequencing runs were used in order to detect Single Nucleotide Polymorphisms (SNPs) in genes related to immune response and gonad differentiation. A total of 866 true SNPs were detected in 140 different contigs representing 262,093 bp as a whole. Only one true SNP was analyzed in each contig. One hundred and thirteen SNPs out of the 140 analyzed were feasible (genotyped), while Ш were polymorphic in a wild population. Transition/transversion ratio (1.354) was similar to that observed in other fish studies. Unbiased gene diversity (He) estimates ranged from 0.060 to 0.510 (mean = 0.351), minimum allele frequency (MAF) from 0.030 to 0.500 (mean = 0.259) and all loci were in Hardy-Weinberg equilibrium after Bonferroni correction. A large number of SNPs (49) were located in the coding region, 33 representing synonymous and 16 non-synonymous changes. Most SNP-containing genes were related to immune response and gonad differentiation processes, and could be candidates for functional changes leading to phenotypic changes. These markers will be useful for population screening to look for adaptive variation in wild and domestic turbot.This study was supported by the Consolider Ingenio Aquagenomics (CSD200700002), the Science and Education Spanish Ministry (AGL2009-11782) and the Xunta de Galicia (09MMA011261PR) projectsS