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Nature-Based Solutions (NbS), a concept introduced in the late 2000s, has developed rapidly during the last years and is now frequently appearing in a broad spectrum of policies developed within the European Union. Its role in marine policies and research programmes is however still limited, but is likely to increase as NbS are adopted as key terminology in both biodiversity strategies and the EU taxonomy for sustainable financing. This will enhance the need for scientific advisory institutions to provide evidence-based advice on potential impacts of various combinations of marine NbS. To facilitate a critical debate about the prospects and pitfalls related to the operationalisation of marine NbS in an EU context, this paper provides an analysis of core definitions, potential categories of marine NBS and a suite of case studies. Coastal waters, shelf and open oceans present multiple options for testing new and scaling up known NbS, which could support both environmental restoration simultaneously with addressing multiple societal challenges, paving the way for a new level of ecosystem-based management. However, as the acceptance of NbS types will depend on ecosystem state and thus history, it will be a significant task to consistently communicate why some solutions may count as a NbS in some areas, while not in others. To conclude, the paper therefore raises a set of research priorities and policy advice aimed at ensuring the successful advice and deployment of marine NBS in support of multiple societal goals.

The Atlantic bluefin tuna (ABFT), Thunnus thynnus (Linnaeus, 1758), is an important commercial species managed as two different stocks, western and eastern Atlantic, with their spawning grounds in the Gulf of Mexico and in the Mediterranean Sea, respectively. The eastern Atlantic stock has been overexploited in the last decades, leading to the application of specific management measures introduced by the International Commission for the Atlantic Tuna (ICCAT). A clear understanding of the genetic structure of ABFT Mediterranean population should be pursued in order to support management decisions. To date the genetic studies on the Mediterranean ABFT, carried out with different molecular markers and sampling procedures, have produced unclear results. Here, we analysed ABFT samples from central and western Mediterranean Sea with mitochondrial sequences and 11 microsatellite loci to investigate, among the others, the area of the Strait of Messina, where environmental conditions seem to support a resident population of ABFT. Furthermore, genetic analyses of mitochondrial sequences were carried out including nucleotide sequences of Adriatic ABFT wild larvae retrieved from GenBank. Among the investigated areas a genetic differentiation was detected between the Strait of Messina and the Tyrrhenian Sea with microsatellite loci according to the exact G test, but not to the Bayesian analyses carried out with STRUCTURE. The analyses with mitochondrial sequences do not reveal any differentiation among sampled areas, however, a highly significant genetic divergence was observed between the Adriatic mitochondrial sequences retrieved from GenBank and the central‐western Mediterranean sequences obtained in the present work. Our results provide some evidence of population structure of Mediterranean ABFT adding pieces to a still unclear picture.

Sustainability of aquaculture is tied to the origin of feed ingredients. In search of sustainable fish meal-free formulations for rainbow trout, we evaluated the effect of Hermetia illucens meal (H) and poultry by-product meal (P), singly (10, 30, and 60% of either H or P) or in combination (10% H + 50% P, H10P50), as partial replacement of vegetable protein (VM) on gut microbiota (GM), inflammatory, and immune biomarkers. Fish fed the mixture H10P50 had the best growth performance. H, P, and especially the combination H10P50 partially restored α-diversity that was negatively affected by VM. Diets did not differ in the Firmicutes:Proteobacteria ratio, although the relative abundance of Gammaproteobacteria was reduced in H and was higher in P and in the fishmeal control. H had higher relative abundance of chitin-degrading Actinomyces and Bacillus, Dorea, and Enterococcus. Actinomyces was also higher in H feed, suggesting feed-chain microbiome transmission. P increased the relative abundance of protein degraders Paeniclostridium and Bacteroidales. IL-1β, IL-10, TGF-β, COX-2, and TCR-β gene expression in the midgut and head kidney and plasma lipopolysaccharide (LPS) revealed that the diets did not compromise the gut barrier function or induce inflammation. H, P, and H10P50 therefore appear valid protein sources in fishmeal-free aquafeeds.

Among the behavioural factors affecting survival of hatchery-reared fish when released into the wild, inefficient foraging is considered to be a major behavioural deficit. The present study investigates the ability of hatchery-produced, pellet-reared dusky grouper Epinephelus marginatus juveniles to recognize and catch live prey (sea bass Dicentrarchus labrax fingerlings 30–80 mm TL) and analyses the effects of a training period on their foraging performance. Juvenile E. marginatus (3+ years, 270–330 mm TL) showed an innate ability to recognize and capture live D. labrax fingerlings as latency time to first attack was very short (

International audience; * To meet the Convention on Biological Diversity's Aichi Target 11 on marine biodiversity protection and Aichi Target 6 on sustainable fisheries by 2020, as well as the Sustainable Development Goal (SDG) 2 on food security and SDG 14 on oceans by 2030, there is an urgent need to rethink how best to reconcile nature conservation and sustainable development. * This paper argues for effective governance to support processes that apply principles of sustainable development and an ecosystem approach to decide about economic activities at sea such as aquaculture. It describes opportunities, benefits and synergies between aquaculture and MPAs as a basis for wider debate. The scope is not a comprehensive analysis of aquaculture and MPAs, but rather to present examples of positive interactions between aquaculture activities and MPAs. The unintended negative consequences are also discussed to present balanced arguments. * This work draws from four workshops held in 2015 and 2016 and used to collect information from about 100 experts representing various sectors and perspectives. * It is recognized that aquaculture is an important activity in terms of sustainable development. It can play a role in providing food security, poverty alleviation and economic resilience, in particular for MPA local communities, and contribute to wild stock enhancement, as an alternative to overfishing and for providing services to the ecosystem. * This study showed that there is a need from both aquaculture and MPA sides for clarity of objectives and willingness for open and extensive dialogue. The paper concludes by describing a number of tools and methods for supporting greater synergies between aquaculture and MPAs. * The results from this work have already helped to build a common understanding between conservation and aquaculture and initiate a rapprochement for increasing synergies.

The presence and distribution of apoptotic cells during thymus development and in adult were studied by in situ end-labelling of fragmented DNA in three temperate species carp (Cyprinus carpio), sea bass (Dicentrarchus labrax) and dusky grouper (Epinephelus marginatus) and in the adult thymus of three Antarctic species belonging to the genus Trematomus spp. During thymus development some few isolated apoptotic cell (AC) firstly appeared in the central–external part of the organ (carp: 5 days ph; sea bass: 35 days ph grouper: 43 days ph). Initially the cells were isolated and then increased in number and aggregated in small groups in the outer-cortical region of the thymus larvae. The high density of apoptotic cells was observed in the junction between cortex and medulla from its appearance (border between cortex and medulla, BCM). ACs decreased in number in juveniles and adult as well as the ACs average diameter. In late juveniles and in adulthood, the apoptosis were restricted to the cortex. In Antarctic species the thymus is highly adapted to low temperature (high vascularisation to effort the circulation of glycoproteins enriched plasma and strongly compact parenchyma). The apoptosis process was more extended (4–7 fold) as compare with the thymus of temperate species, even if the distribution of ACs was similar in all examined species. Data suggested a common process of T lymphocyte negative-selection in BCM of thymus during the ontogeny. The selection process seems to be still active in adult polar fish, but restricted mainly in the cortex zone.

A genetic survey of Barbus spp. populations in the Marches Region (Adriatic River basins), central Italy, was carried out using mitochondrial and nuclear markers (partial D-loop, cyt b sequences and microsatellite loci) in order to ascertain their systematic position and to address their genetic structure which is key to conservation action planning. Analyses were conducted on sequences obtained from 91 individuals collected from eight sampling sites in five different rivers, from two specimens provided by the Ichthyological Centre of Rome and mitochondrial sequences of Barbus spp. retrieved from GenBank. Presumptive classification based on external morphological characters was not confirmed by genetic analysis, by means of which all specimens collected in the Marches Region were ascribed to Barbus plebejus. Genetic diversity values (h and π) of sampling groups were all different from 0 except the one sample collected from the upper reaches of the River Tenna, above a hydroelectric dam. Population connectivity and colonization patterns of the studied area were inferred from an analysis of molecular variance distribution and evolutionary relationships among haplotypes. The results point to different levels of isolation among sampling groups due to ecological and anthropogenic factors and the effect of an artificial barrier on genetic variability and conservation status of the population. Finally, this study confirms the uncertainty associated with systematic classification of Barbus spp. based on morphological characters due to the phenotypic plasticity of the species.

Aquaculture is an increasingly important food-producing sector, providing protein for human consumption. However, marine aquaculture often struggles for space due to the crowded nature of human activities in many marine coastal areas, and because of limited attention from spatial planning managers. Here, we assess the need for coastal spatial planning, emphasising the establishment of suitable areas for the development of marine aquaculture, termed Allocated Zones for Aquaculture (AZAs), in which aquaculture has secured use and priority over other activities, and where potential adverse environmental impacts and negative interactions with other users are minimised or avoided. We review existing examples of marine aquaculture spatial development worldwide and discuss the proper use of site selection in relation to different legal and regulatory requirements. National or regional authorities in charge of coastal zone management should carry out spatial planning defining optimal sites for aquaculture to promote development of sustainable marine aquaculture and avoid conflict with other users, following a participatory approach and adhering to the principles of ecosystem-based management., peer-reviewed

Individual variation in stress physiology and behaviour has been previously reported in several fish species. As seen in other vertebrates, existence of stress coping styles seems to be reflected by the presence of individual variation. Aggressive behaviour, amongst others, is one of the most commonly used parameters to characterize coping styles. However, not all fish species exhibit aggressive behaviour, such as the flatfish Senegalese sole, Solea senegalensis (Kaup, 1858). Therefore, the goal of this study was to determine the magnitude of individual variation in behavioural parameters other than aggression (feeding motivation and activity during stress) as well as in growth and stress response in Senegalese sole. The relationship between these variables was investigated to determine whether they could be used as indicators of coping styles. Thirty-six juvenile fish (9.9 2.2 g) were individually housed for 73 days. Feeding motivation, measured as the time (in s) taken by each fish to react to feed, was determined on days 10, 17, 24 and 31. Blood samples for plasma cortisol were collected on days 51 and 71 for determination of undisturbed and stress levels, respectively. The stress test consisted of holding each fish individually in a net, outside the water, for 3 min. Duration of escape attempts, i.e. the time taken by each fish to stop struggling (in an attempt to escape) in the net, was quantified. The results showed a pronounced individual variation in both control (CV = 54%) and acute stress (CV = 71%) cortisol levels. Senegalese sole also exhibited high coefficient of variation in the behavioural parameters: 75% in feeding latency and 96% in duration of escape attempts. Growth (RGR = 1.17 0.38) showed to be the parameter with lower variation of only 32% and was not correlated with any of the measured parameters. A significant correlation between undisturbed cortisol levels and duration of escape attempts was found. Undisturbed cortisol levels (8.08 4.36 ng/ml) were negatively correlated with duration of escape attempts (P = 0.009, rs =0.503). Correlations between plasma cortisol levels after stress (398.45 282.67 ng/ml) and the behavioural parameters were not found. The observed individual variation in behaviour and stress physiology as well as their relationship suggests the existence of coping styles in Senegalese sole where proactive individuals exhibit shorter feeding latency, higher duration of escape attempts and lower undisturbed cortisol levels than passive individuals.

The first attempts to reproduce dusky grouper (Epinephelus marginatus, Lowe 1834) under controlled conditions started in 1995, but the egg and larvae quality was very low. Mass production is still encountering many difficulties, mainly concentrated in the larval period when very high mortality rates are observed, confirming what has been observed in the rearing of other grouper species. The main bottlenecks have been identified as the difficulty to properly nourish the larvae, stress shock syndrome, and the high deformation rates. We analysed 633 dusky grouper larvae and juveniles (0.2–7.2 cm total length, TL), sampled during two larval rearing cycles carried out in 2001 and 2002 in Italy. The specimens at different development stages were stained in toto for bone and cartilage and examined for skeletal anomalies during dusky grouper ontogenesis. The incidence of anomalies in groupers hatched from the same egg batch but reared using two different methods (green waters and semi-intensive rearing) and three stocking densities (8, 16 and 28 larvae/l) was compared, with a view to providing tools for identifying the most appropriate larval rearing method in order to at least limit the onset of skeletal anomalies. Our results suggest that during development no particular skeletal anomaly patterns (or fate) can be clearly identified as a high variability was observed in malformation typologies and the regions affected. No significant differences in the morphological quality between groupers reared using semi-intensive (LV02 lot) and green water (GW02-01 lot) methodologies were observed, whilst groupers reared at the highest stocking density (28 larvae/l) showed the highest frequency of deformed individuals (75.8%), the highest malformation charge (average of 5.5 anomalies per deformed individual), the largest range of anomaly typologies (38), and the highest incidence of individuals with at least one severe anomaly (30.9%). Whilst in green waters no evident effects of larvae density were observed on survival rates, the survival rate in large volume reared individuals (17.5%) was considerably higher with respect to those reared in green waters (0.2%) at 7–8 larvae/l. This indicates that the semi-intensive methodology should be considered more effective in enhancing the survival rate of dusky grouper larvae.

The dusky grouper (Epinephelus marginatus) is an endangered marine fish for which a restocking program has been undertaken in the Mediterranean Sea. Genetic variation at six microsatellite loci was assayed to help plan preservation of genetic diversity after release of hatchery-reared juveniles. We estimated relatedness and effective population size within two groups of captive broodstock, assessed genetic variability of these broodstock and F1 juveniles compared to natural populations, and investigated genetic relationships between hatchery-reared and wild specimens inhabiting the release site. On average, no relatedness was found within captive broodstock and a potentially high number of effective breeders was assessed (N b = 86.9; CI 95% = 46.0–451.2). However, very few breeders (N = 7) spawned successfully, and a positive degree of relatedness (r W = 0.0297) was found among this particular subset of individuals. Genetic variability parameters were reduced for the broodstock and F1 juveniles, although ...

The impact of fish meal replacement with plant proteins on fish health is still poorly investigated. This study evaluates the effects of several diets containing graded levels of a mixture of gluten meal, soybean meal and soy protein concentrate on some immune parameters of turbot juveniles. The feeding trial was carried out at the aquaculture facilities of the Veterinary University of Bologna (Cesenatico, Italy). Turbot juveniles (9.7kept at 18in 500 l recirculating tanks (in triplicate) were fed four isonitrogenous (51%) and isolipidic diets (16%) (Skretting ARC, Stavanger, NO) for 9 weeks: a reference diet of 50% fish meal (FM50), and three diets containing 35%, 20% and 5% fish meal (FM35; FM20; FM5), with a proportional amount of vegetable ingredients. At the end of the trial, blood samples and spleen tissues were collected and analyzed for serum lysozyme concentration (lysoplate assay), differential leukocyte count (blood smears/May-Grunwald Giemsa) and quantification of splenic melano-macrophage centres (MMCs) (histology/image analyzer). One-way ANOVA and c2-test were applied respectively to humoral and cellular immune parameters to test the effects of dietary treatment. Plant protein diets induced significant changes in several immune parameters in turbot juveniles. Serum lysozyme was higher in turbot fed on FM35 and FM20 diets. The percentage of circulating phagocytes, mainly neutrophils, and splenic MMCs, increased, especially in fish fed FM5 and FM20 diets. No difference was observed in lymphocyte population. The increase in blood phagocytes could be responsible for the rise in serum lysozyme and interpretable as a stress or inflammatory response. Conversely, the lower lysozyme level measured in turbot fed FM5 diet in the face of a higher number of circulating phagocytes could be interpreted as a functional impairment of these cells. The increase in MMCs may be associated with a nutritional deficit, which is more severe in fish fed FM5. Long-term feeding trials are needed in order to gain further insight into the impact of dietary plant proteins on immune function and health in turbot.

The reproductive biology of the Mediterranean razor fish Xyrichthys novacula was investigated by demographic data and histological analysis of the female, intersexual and male gonads. Specimens were collected by bottom trawl on a monthly basis between June 2000 and July 2001 in a sandy bay in southern Thyrrenian. Gonad histology confirmed that the Mediterranean razor fish is a monandric, protogynous hermaphrodite. Females reached first sexual maturity at 100 mm (L T ) and the estimated mean LT at first maturity (L 50 ) was 125 mm. Females exhibited asynchronous ovarian development and multiple ovulations occurred over the spawning period. Vitellogenesis started in early May and spawning occurred from late May until late September. Sexual transition involved a large-scale atresia of all oocyte stages and a massive degeneration of ovarian tissue followed by primordial germ cells proliferation. Sex change began at spawning time (June) but transitional individuals tended to cluster at the end of the reproductive period (September). They accounted for 17.1% of the population sampled and were found in a broad size range (105-150 mm L T ).

Captive-reared dusky grouper were induced to ovulate using a sustained-release delivery system (implant) loaded with gonadotropin-releasing hormone agonist [D-Ala 6 , Pro 9 , NEt]-GnRH (GnRHa). Thirteen females were implanted at doses ranging from 30.5 to 68.3 A gk g 1 during three experiments between late June and early September. Of the injected females, 85% responded positively to the GnRHa implant and ovulated between 60 and 238 h after treatment, whereas none of control fish showed any sign of maturation. No spontaneous spawning was observed, and the eggs were manually removed from the females using gentle abdominal pressure. The mean number of ovulations per fish was 3.8, with a maximum of nine for one female. Overall, a total of 42 ovulations were obtained, resulting in the production of more than 5 million eggs. The average relative fecundity was 118.3F16.010 3 eggs kg 1 BW, with a maximum of 202.210 3 eggs kg 1 BW. Mean percentage fertilisation and hatching were 48.2% and 52.2%, respectively. The results demonstrate that GnRHa administration via controlled delivery systems is an effective method for producing good quality eggs in captive dusky grouper. D 2003 Elsevier Science B.V. All rights reserved.

Background Water temperature greatly influences the physiology and behaviour of teleost fish as other aquatic organisms. While fish are able to cope with seasonal temperature variations, thermal excursions outside their normal thermal range might exceed their ability to respond leading to severe diseases and death. Profound differences exist in thermal tolerance across fish species living in the same geographical areas, promoting for investigating the molecular mechanisms involved in susceptibility and resistance to low and high temperatures toward a better understanding of adaptation to environmental challenges. The gilthead sea bream, Sparus aurata, is particularly sensitive to cold and the prolonged exposure to low temperatures may lead to the "winter disease", a metabolic disorder that significantly affects the aquaculture productions along the Northern Mediterranean coasts during winter-spring season. While sea bream susceptibility to low temperatures has been extensively investigated, the cascade of molecular events under such stressful condition is not fully elucidated. Results In the present study two groups of wild sea bream were exposed for 21 days to two temperature regimes: 16 ± 0.3°C (control group) and 6.8 ± 0.3°C (cold-exposed group) and DNA microarray analysis of liver transcriptome was carried out at different time points during cold exposure. A large set of genes was found to be differentially expressed upon cold-exposure with increasingly relevant effects being observed after three weeks at low temperature. All major known responses to cold (i.e. anti-oxidant response, increased mitochondrial function, membrane compositional changes) were found to be conserved in the gilthead sea bream, while, evidence for a key role of unfolded protein response (UPR) to endoplasmic reticulum (ER) stress, during short- and long-term exposure to cold is reported here for the first time. Conclusions Transcriptome data suggest a scenario where oxidative stress, altered lipid metabolism, ATP depletion and protein denaturation converge to induce ER stress. The resulting UPR activation further promotes conditions for cell damage, and the inability to resolve ER stress leads to severe liver dysfunction and potentially to death. Electronic supplementary material The online version of this article (doi:10.1186/1471-2164-15-765) contains supplementary material, which is available to authorized users.

The EU Marine Strategy Framework Directive 2008/56/EC (MSFD) defines a framework for Community actions in the field of marine environmental policy in order to achieve and/or maintain the Good Environmental Status (GES) of the European seas by 2020. Microbial assemblages (from viruses to microbial-sized metazoa) provide a major contribution to global biodiversity and play a crucial role in the functioning of marine ecosystems, but are largely ignored by the MSFD. Prokaryotes are only seen as "microbial pathogens", without defining their role in GES indicators. However, structural or functional prokaryotic variables (abundance, biodiversity and metabolism) can be easily incorporated into several MSFD Descriptors (i.e. D1. Biodiversity, D4. Food webs, D5. Eutrophication, D8. Contaminants and D9. Contaminants in seafood) with beneficial effects. The present review provides a critical analysis of the current MSFD descriptors and illustrates the reliability and advantages of the potential incorporation of some prokaryotic variables within the set of indicators of marine environmental quality. Following a cost/benefit analysis against scientific and economic criteria, we conclude that marine microbial components, and particularly prokaryotes, are highly effective for detecting the effects of anthropogenic pressures on marine environments and for assessing changes in the environmental health status. We thus recommend the inclusion of these components in future implementations of the MSFD.

Demographic data and gonad histology confirmed that the dusky grouper Epinephelus marginatus is a protogynous hermaphrodite that follows a monandric pathway to sexual development. Females reached first sexual maturity at 36.7 cm L S and estimated mean length at first maturity (L 50 ) was 43.8 cm L S for females and 81.3 cm L S for males. Adult sex ratios during the reproductive period were c. 3.5: 1 females to males. Females exhibited group-synchronous ovarian development and multiple ovulation occurred over the spawning period. Gonads were ripe from early May and spawning occurred from June until early September. The size of ripe testes (0.6% W) indicated strong oligospermy and suggested a mating system with no sperm competition. Sexual transition was protogynous involving regression of ovarian tissue and proliferation of testicular tissue in the gonads. Transitional individuals occurred from May through November and accounted for 9% of sampled adult population. Sex change occurred in fish 69-93 cm (L S ) long and the size distributions of males and females overlapped over 27% of the L S range Special zones were recognized as gathering areas for sexually mature dusky groupers during the reproductive period.

A cytogenetic analysis was carried out on specimens of Epinephelus marginatus (Lowe, 1834) from three localities in the Mediterranean Sea, to deepen knowledge of the chromosome complement of the species and identify any possible population-specific cytogenetic markers. All specimens had a 2n = 48 acrocentric karyotype with two Ag-, chromomycin A3- and C-positive NORs (nucleolar organizer regions) in the subcentromeric region of the smallest chromosome pair. The constitutive heterochromatin was distributed centromerically. Except for NORs, neither eu- nor heterochromatin show fluorescence after fluorochrome staining, i.e. there is no localized increase of AT- or GC-rich DNA. In all populations, the (TTAGGG)n telomeric sequences are restricted to the telomeres, and the 18S and the 5S rDNA clusters are located on different chromosome pairs. In specimens from Sardinia, additional signals after C-banding, Ag-staining and FISH (fluorescence in situ hybridization) with 18S rDNA can be observed in the telomeric region of one or two large-sized chromosomes, classified as No. 2. This suggests that additional and variable NORs could be detected in the species in addition to those on the genus-specific, NOR-bearing Chromosome Pair 24.

Wild Seriola dumerilii were collected in the South Mediterranean Sea during the 1990, 1991 and 1992 spawning seasons. Macroscopic and cytological characteristics of ovary and testis were analysed. Depending on the presence and the number of oocytes at different stages, a five point maturity scale was proposed for ovarian maturity. Nine maturity stages of the oocytes are described. Oocyte size-frequency distribution has shown a group synchronous ovarian development type. The testes have a typical lobular-type structure. Depending on lobules and sperm duct development and on the abundance of germ cells, testis maturity was classified in four stages. Maturity rate according to age and size was also determined. 100% of 1-year-old fish show immature gonads. The proportion of females with mature ovaries was 0, 12.5, 84.6 and 100% at the age of 2, 3, 4 and 5 years respectively. The proportion of males with mature testes was 14.3, 40, 80 and 100 at the same age. The median standard length at which 50% of the fish attained maturity is 109 and 113 cm SL in males and females respectively.

Summary Hatche and wild sea bass (Dicentrarchus labrax) fry were transferred from sea water to low salinities and freshwater (FW) usiny three different operational protocols: 1) direct exposure to ow salimues and FW (LC 50 test, ST 50 test); 2) quick acclimation to FW (48h); 3) slow acclimation to FW (17d). Direct exposure to freshwater led to e death of all fry. Comete survival was observed after direct transfer to salinities ≥ 9 ppt, both in wild and hatchery fry. Eatchery fry tolerate direct transfer to low salinities (LC 50=4.393 ppt) better than wild fry (LC 50 = 3.215 ppt). Slow acclimation rotocol leads to higher survival rates (nearly 90 %) than does the quick procedure. In 48h and 17d accimation trials wild fry tolerate FW better tlan hatchery fry. Hypotheses to explain observed differences in wild and hatchery tolerances to low salinities and FW are discussed.

The development of the fin skeleton has been studied in both the wild and hatchery sea bass, Dicentrarchus labrax L., larvae, in order to identify and define morphofunctional criteria for larval quality assessments. The cartilaginous and bony skeletal elements of the larvae were studied and standard length and notochord flexure recorded. Fin anomalies were present in reared larvae, whereas they were rare or absent in specimens from the wild. These anomalies were recognizable from the very beginning of skeletal development, indicating that environmental, genetic and broodstock management factors influence even the very early stages of larval development.

The aim of this study was to identify possible metabolic molecular indicators of chronic stress in gilthead sea bream Sparus aurata . Two potential stressful conditions were tested: repeated handling and crowding at high stocking density. Gilthead seabream kept under optimized rearing conditions were used as control fish. Cortisol was measured as primary stress indicator and the liver proteome of stressed fish was compared to that of control fish using comparative proteomics. Plasma cortisol levels in sea bream repeatedly handled and crowded at high stocking density were significantly higher than in undisturbed control fish. A total of 560 spots were detected and the statistical analysis revealed a differential expression in about 50% of all detected proteins. Spots with greater than 2-fold or lower than 0.5-fold changes were identified by liquid chromatography–tandem mass spectrometry (LC–MS/MS). Proteins like fatty acid binding protein (lipid transport and antioxidant role), heat shock cognate protein (chaperoning), calmodulin (Ca 2+ signaling), mitochondrial porine — voltage-dependent anion channel (lipid oxidation), glutamine synthetase (ammonia metabolism), cofilin and beta-tubulin (cytoskeleton), hemoglobin and several other proteins involved in carbohydrate metabolism (triose-phosphate isomerase, pyruvate dehydrogenase, glyceraldehyde 3-phosphate dehydrogenase, alfa-enolase) were differentially expressed in fish under chronic stress. Some of these proteins may be used in the future as chronic stress and/or part of a panel of welfare biomarkers, after validation studies using RT-PCR and ELISA assays.

A pilot experiment of stock enhancement based on the release of hatchery-reared juveniles of dusky grouper, Epinephelus marginatus, was carried out at two artificial reefs located 0.9 km offshore the south-western Sicilian coast (Central Mediterranean). The fish assemblages inhabiting the reefs were characterized using underwater visual census (UVC) surveys. Despite some differences in fish species richness, diversity and abundance, both artificial reefs provide suitable feeding resources and shelter opportunities for the settlement of dusky grouper juveniles. A total of 95 dusky grouper juveniles, which were 3 and 4 years old, were released. No stress-related effects on fish swimming or behaviour were observed during the step by step transportation to the sea-bottom at 20 m depth. After release, several groupers showed very low reactivity; freezing and schooling were the most common behaviours. The sighting rate after one month of UVC surveys was 15.7% and 20% at the two artificial reefs. No effect of fish size at release on grouper survival was observed. Some groupers were not recorded on the pyramid of release but elsewhere in the same artificial reef, which demonstrated their ability to move around the reef. The extent of dispersal of the released juveniles largely exceeded the area encompassed by the reefs, which was shown by recaptures of tagged groupers by local fishermen up to 13 km from the release site. This result provides a first insight into the potential of stock enhancement of hatchery reared dusky grouper juveniles for marine ranching and conservation purposes., Un experimento de repoblación, basado en la liberación de juveniles de mero Epinephelus marginatus cultivados en criadero, fue realizado en dos arrecifes artificiales situados 0.9 kilómetros fuera de las costas sicilianas sur-occidentales (Mediterráneo Central). La ictiofauna de los arrecifes fue caracterizada a través de inmersiones submarinas, utilizando la técnica de censo visual (UVC). A pesar de algunas diferencias en número de especies, diversidad y abundancia, ambos arrecifes artificiales proporcionaron recursos alimenticios y refugio adecuados para el establecimiento de juveniles de mero. En total se liberaron 95 meros juveniles de 3 y 4 años de edad. Ningún efecto relacionado al estrés fue observado en el comportamiento de natación de los peces durante el gradual transporte hasta 20 m de profundidad. Después de la liberación, varios meros mostraron una reactividad muy baja, siendo el comportamiento más común quedarse inmóviles y agregados. Después de un mes de censos visuales, las tasas de avistamientos obtenidas en los arrecifes artificiales fueron de 15.7% y 20%. No se observó relación entre el tamaño en el momento de la liberación y la supervivencia. Algunos peces no fueron observados cerca del punto de liberación sino en otra parte del mismo arrecife artificial, demostrando así su capacidad de moverse por los alrededores. Según lo indicado por los pescadores locales que recobraron meros marcados hasta 13 kilómetros del punto de liberación, el grado de la dispersión de los juveniles liberados superó ampliamente el área abarcada por los arrecifes. Este resultado proporcionan la primera evaluación del potencial de los juveniles de E. marginatus cultivados en criadero para actividades de cría en mar y conservación.

A pilot experiment of stock enhancement based on the release of hatchery-reared juveniles of dusky grouper, Epinephelus marginatus, was carried out at two artificial reefs located 0.9 km offshore the south-western Sicilian coast (Central Mediterranean). The fish assemblages inhabiting the reefs were characterized using underwater visual census (UVC) surveys. Despite some differences in fish species richness, diversity and abundance, both artificial reefs provide suitable feeding resources and shelter opportunities for the settlement of dusky grouper juveniles. A total of 95 dusky grouper juveniles, which were 3 and 4 years old, were released. No stress-related effects on fish swimming or behaviour were observed during the step by step transportation to the sea-bottom at 20 m depth. After release, several groupers showed very low reactivity; freezing and schooling were the most common behaviours. The sighting rate after one month of UVC surveys was 15.7% and 20% at the two artificial reefs. No effect of fish size at release on grouper survival was observed. Some groupers were not recorded on the pyramid of release but elsewhere in the same artificial reef, which demonstrated their ability to move around the reef. The extent of dispersal of the released juveniles largely exceeded the area encompassed by the reefs, which was shown by recaptures of tagged groupers by local fishermen up to 13 km from the release site. This result provides a first insight into the potential of stock enhancement of hatchery reared dusky grouper juveniles for marine ranching and conservation purposes. Un experimento de repoblación, basado en la liberación de juveniles de mero Epinephelus marginatus cultivados en criadero, fue realizado en dos arrecifes artificiales situados 0.9 kilómetros fuera de las costas sicilianas sur-occidentales (Mediterráneo Central). La ictiofauna de los arrecifes fue caracterizada a través de inmersiones submarinas, utilizando la técnica de censo visual (UVC). A pesar de algunas diferencias en número de especies, diversidad y abundancia, ambos arrecifes artificiales proporcionaron recursos alimenticios y refugio adecuados para el establecimiento de juveniles de mero. En total se liberaron 95 meros juveniles de 3 y 4 años de edad. Ningún efecto relacionado al estrés fue observado en el comportamiento de natación de los peces durante el gradual transporte hasta 20 m de profundidad. Después de la liberación, varios meros mostraron una reactividad muy baja, siendo el comportamiento más común quedarse inmóviles y agregados. Después de un mes de censos visuales, las tasas de avistamientos obtenidas en los arrecifes artificiales fueron de 15.7% y 20%. No se observó relación entre el tamaño en el momento de la liberación y la supervivencia. Algunos peces no fueron observados cerca del punto de liberación sino en otra parte del mismo arrecife artificial, demostrando así su capacidad de moverse por los alrededores. Según lo indicado por los pescadores locales que recobraron meros marcados hasta 13 kilómetros del punto de liberación, el grado de la dispersión de los juveniles liberados superó ampliamente el área abarcada por los arrecifes. Este resultado proporcionan la primera evaluación del potencial de los juveniles de E. marginatus cultivados en criadero para actividades de cría en mar y conservación.

Mediterranean amberjacks, Seriola dumerilii Risso, were caught off the Pelagie Islands, in the south Mediterranean Sea, between May 1997 and June 1999. Fish blood was sampled, and gonads were collected at 10-day intervals throughout the spawning period and at monthly intervals during the resting period. Concentrations of plasma estradiol-17s (E2), testosterone (T), 17,20s-dihydroxy-4-pregnen-3-one (DHP) and vitellogenin (Vtg) in females; and plasma T and 11-ketoT (11-KT) in males were correlated with changes in gonadal development. The first females that had already ovulated (F5) were found in late May. Most mature females (F4) were caught in June. Post-spawned females (F6) were found from late July until September. Estradiol-17s was at baseline levels (

Modifications of the arbitrarily primed polymerase chain reaction assay (i.e. a low annealing temperature and a very slow increase in the temperature during the elongation steps during the amplification cycles) allowed it to be used with the AT-rich Plasmodium falciparum DNA. The analysis of the products by polyacrylamide-urea gels, after silver staining, resulted in high resolution and sensitivity. Eighteen single and six combined pairs of arbitrary primers were tested. Two produced polymorphic patterns complex enough to differentiate between close Colombian isolates in a single assay. This method may be useful in studying the distribution and migration of strains in endemic areas, and for identifying intralaboratory cross-contamination of cultures.

Gravid females of the nematode Philometra lateolabracis (Yamaguti, 1935), a parasite of gonads of marine perciform fishes, were found in wild and cultured dusky grouper Epinephelus marginatus (Lowe) from waters near the Balear Islands (Spain, Mediterranean Sea) and Sicily (Italy, Thyrrenean Sea), and in the greater amberjack Seriola dumerili (Risso) in Croatia (south-eastern Adriatic Sea). In wild E. marginatus in Spain, the overall prevalence was 21% and the intensity of infection 1 nematode per fish. The nematodes are briefly described and illustrated. The species Sanguinofilaria jordanoi López-Neyra, 1951, described from the ovary of Epinephelus gigas Brünich from Morocco, is synonymized with P. lateolabracis. This is the first documented record of P. lateolabracis from fishes of the Mediterranean region and its finding in S. dumerili represents a new host record. The possible importance of this pathogenic parasite for cultures of marine perciform fishes in the region is stressed.