Environmental Conditioning of Skeletal Anomalies Typology and Frequency in Gilthead Seabream (Sparus aurata L., 1758) Juveniles

In this paper, 981 reared juveniles of gilthead seabream (Sparus aurata) were analysed, 721 of which were from a commercial hatchery located in Northern Italy (Venice, Italy) and 260 from the Hellenic Center for Marine Research (Crete, Greece). These individuals were from 4 different egg batches, for a total of 10 different lots. Each egg batch was split into two lots after hatching, and reared with two different methodologies: intensive and semi-intensive. All fish were subjected to processing for skeletal anomaly and meristic count analysis. The aims involved: (1) quantitatively and qualitatively analyzing whether differences in skeletal elements arise between siblings and, if so, what they are; (2) investigating if any skeletal bone tissue/ossification is specifically affected by changing environmental rearing conditions; and (3) contributing to the identification of the best practices for gilthead seabream larval rearing in order to lower the deformity rates, without selections. The results obtained in this study highlighted that: i) in all the semi-intensive lots, the bones having intramembranous ossification showed a consistently lower incidence of anomalies; ii) the same clear pattern was not observed in the skeletal elements whose ossification process requires a cartilaginous precursor. It is thus possible to ameliorate the morphological quality (by reducing the incidence of severe skeletal anomalies and the variability in meristic counts of dermal bones) of reared seabream juveniles by lowering the stocking densities (maximum 16 larvae/L) and increasing the volume of the hatchery rearing tanks (minimum 40 m³). Feeding larvae with a wide variety of live (wild) preys seems further to improve juvenile skeletal quality. Additionally, analysis of the morphological quality of juveniles reared under two different semi-intensive conditions, Mesocosm and Large Volumes, highlighted a somewhat greater capacity of Large Volumes to significantly augment the gap with siblings reared in intensive (conventional) modality.     

Effect of rearing techniques on skeletal deformities and osteological development in red porgy Pagrus pagrus (Linnaeus, 1758) larvae

Red porgy is a candidate species for marine aquaculture diversification. The objective of the present study was to describe the osteological development in this species and the occurrence of skeletal deformities in relation to the intensification of the rearing system. Fish samples were periodically collected along the development from hatching to juveniles (95 days after hatching). Osteological development and the presence of skeleton abnormalities were evaluated. Larvae reared under S‐IS showed a better growth in terms of total length in comparison with IS reared ones. Regarding to osteological development for red porgy, this was similar between fish from both culture systems, but differing in timing of apparition and ossification of skeletal elements. X‐ray studies revealed a high number of fish with skeletal deformities (Semi‐intensive: 38.8%; Intensive: 46.5%), but no significant effect of the rearing technique on the incidence of deformities such as lordosis or fused vertebrae was found. However, cranial abnormalities and kyphosis incidences were significantly higher in intensive system cultured red porgy. These results, suggest a relationship among rearing technique, osteological development and the apparition of certain deformities.     

Occurrence of skeletal deformities and osteological development in red porgy Pagrus pagrus larvae cultured under different rearing techniques

The present study describes the osteological development and the occurrence of skeletal deformities in red porgy Pagrus pagrus larvae in relation to the intensification of the rearing system. Eggs obtained from natural spawning were cultured under two different rearing systems: intensive (100 eggs l^?1) in 2000 l and semi‐intensive (mesocosm) system (5 eggs l^?1) in 40 000 l conico‐cylindrical tanks. Fish samples were periodically collected along the development from hatching to juveniles at 95 days post hatching (dph). Osteological development, meristic counts and the presence of skeletal deformities were evaluated. Despite the external appearance of the juveniles being similar to wild standards, X‐ray studies revealed a high number of fish (semi‐intensive: 37·8%; intensive: 45·5%) with skeletal deformities. Regardless of the rearing system, no significant interaction was found between the per cent of the most common deformities, axial deviations (lordosis and presence of fused vertebrae). Cranial deformities and kyphosis incidences, however, were significantly higher in intensively cultured P. pagrus. Also, the fused vertebrae in these fish were located mainly in the caudal area instead of pre‐haemal area for semi‐intensively reared P. pagrus. Moreover, a significant interaction was found between the total number of vertebrae and the type of rearing system used; fish from the intensive system showing a higher number of fish with an extra vertebrae (10 abdominal + 15 caudal). Present results suggest a relationship among feeding sequence, osteological development and deformity incidence and location in P. pagrus larvae.     

Gonadal maturation in the blackspot seabream Pagellus bogaraveo: a comparison between a farmed and a wild broodstock

The blackspot seabream Pagellus bogaraveo (Brünnich, 1768) has been regarded as a possible alternative to traditionally cultured Mediterranean species such as seabream and seabass, due to its high market value and good adaptation to captivity. Broodstock establishment and management represent the first step towards reliable production of eggs and fry, which is required to develop aquaculture of this new species. Two different broodstocks were tested for gonadal maturation and spawning, one constituting of wild fish caught as juveniles and reared in tanks until sexual maturity (4 years), and one assembled from wild adult fish caught during or just before the reproductive season. All fish were maintained under the same rearing conditions and fed the same diet. Gonadal stripping and biopsies were performed weekly to monitor maturation in both males and females. Ovarian samples were staged for maturity on the basis of follicular diameter and migration of germinal vesicle. Sperm samples were tested for density (number of spermatozoa ml⁻¹) and motility. The fish reared in captivity reached ovarian maturity during the breeding season of the wild stock. Eggs were obtained by stripping from both farmed and wild specimens, but appeared degenerated as a result of being retained too long in the ovarian cavity due to the absence of spontaneous spawning. Spermiation was prolonged in the farmed fish, but appeared to be blocked in the wild breeders after first sampling. However, the sperm was very viscous and the motile spermatozoa did not exceed 10%.     

Use of meristic counts and skeletal anomalies to assess developmental plasticity of farmed rainbow trout (Oncorhynchus mykiss,Walbaum 1792): a preliminary study

Rainbow trout is the longest‐standing intensively‐reared species in Western countries, where it has been subjected by farmers to selection for productive traits for more than a century. One interesting aspect in this context is to try and understand if prolonged artificial selection has in some way altered the phenotypic plasticity of this species. In this preliminary study, we characterized meristic counts and skeletal abnormalities in two different (Italian and French) rainbow trout strains, reared during on‐growing following different approaches (intensive and semi‐intensive), in order to analyze whether there are any differences in the skeletal elements and if so, what. Despite the different genetic origin and rearing methodologies to which the lots analyzed were subjected, the occurrences of malformed and seriously malformed individuals were not significantly different, nor were the mean number of anomalies per specimen. The body region most affected by deformities was the caudal vertebral column in both lots, although the French lot was also characterized by an high occurrence of spinal anomalies in the haemal region. Among the meristic counts, only the number of anal and dorsal rays showed statistically different median values between the two lots.     

Changes in the soluble bone proteome of reared white seabream (Diplodus sargus) with skeletal deformities

One of the main constrains for commercial aquaculture production of white seabream (Diplodus sargus) is the high incidence of skeletal malformations in reared fish. The purpose of this study was to obtain a better understanding of the mechanisms involved in the development of these types of skeletal malformations by comparative proteomic analysis of the vertebral column of normal and deformed fish using 2DE for protein separation and MS for protein identification. We observed a 3.2 and 3.4-fold increase in the expression of two tropomyosin isoforms, one of which (tropomyosin-4) is essential for the motility and polarization cycles of osteoclasts. Furthermore, a 1.6, 1.7 and 1.8-fold increase in three parvalbumin spots was detected, suggesting a cellular response to increased intracellular Ca²⁺ levels. These results can be interpreted as signs of increased cellular activity in the bone of white seabream with skeletal deformities coupled to a higher degree of calcium mobilization, which elicits further studies into the use of these proteins as indicators of skeletal metabolic state.     

Skeletal Anomaly Monitoring in Rainbow Trout (Oncorhynchus mykiss,Walbaum 1792) Reared under Different Conditions

The incidence of skeletal anomalies could be used as an indicator of the “quality” of rearing conditions as these anomalies are thought to result from the inability of homeostatic mechanisms to compensate for environmentally-induced stress and/or altered genetic factors. Identification of rearing conditions that lower the rate of anomalies can be an important step toward profitable aquaculture as malformed market-size fish have to be discarded, thus reducing fish farmers’ profits. In this study, the occurrence of skeletal anomalies in adult rainbow trout grown under intensive and organic conditions was monitored. As organic aquaculture animal production is in its early stages, organic broodstock is not available in sufficient quantities. Non-organic juveniles could, therefore, be used for on-growing purposes in organic aquaculture production cycle. Thus, the adult fish analysed in this study experienced intensive conditions during juvenile rearing. Significant differences in the pattern of anomalies were detected between organically and intensively-ongrown specimens, although the occurrence of severe, commercially important anomalies, affecting 2–12.5% of individuals, was comparable in the two systems. Thus, organic aquaculture needs to be improved in order to significantly reduce the incidence of severe anomalies in rainbow trout.     

Studies on the appearance of skeletal anomalies in red porgy: effect of culture intensiveness,feeding habits and nutritional quality of live preys

Despite the great interest of red porgy as a new species for Mediterranean aquaculture, its commercial production is constrained by the high incidence of skeletal deformities occurring in this species under culture conditions. Several studies have been conducted to better understand the origin of these anomalies in this species, using different system intensiveness, rotifers enrichment products or rotifers docosahexaenoic acid content. The first study showed that culture intensification increased the number of fish with an extra vertebrae, what was probably related to the different nutritional quality of live preys employed in each treatment, since water temperature, salinity and genetic background were identical for the different batches of fish studied. Total incidence of skeletal abnormalities was higher in the intensive system, particularly cranial abnormalities and kyphosis in the cephalic vertebrae. In both rearing systems the most common skeletal anomalies were vertebral column disorders, lordosis and fused vertebrae, their localization along the column being affected by the culture intensiveness. Rotifer enrichment, predominantly its docosahexaenoic acid content significantly affected deformities occurrence. A marked positive effect of rotifer docosahexaenoic acid content was found on larval survival. X‐ray studies denoted elevated levels of bone abnormalities associated, in both trials, to low docosahexaenoic acid content in live preys. Among different anomalies, the presence of fused vertebrae was the most frequent deformity for both rearing trials. A 50% reduction in the number of deformed fish for each type of deformity was obtained when the larvae were fed higher docosahexaenoic acid levels, denoting the important role of this fatty acid in bone development. Further studies are needed to elucidate the importance of essential fatty acids on the development of bone deformities in fish, since the functions of these fatty acids differ among them and can lead to very different effects in fish metabolism, including bone formation.     

Assessment of lipid and essential fatty acids requirements of black seabream (Spondyliosoma cantharus) by comparison of lipid composition in muscle and liver of wild and captive adult fish

The primary aim of the present study was to compare the contents of total lipid, lipid classes and their associated fatty acids in muscle and liver of wild and one-year captive black seabream (Spondyliosoma cantharus) adults, in order to elucidate the lipid and fatty acids requirements of this fish species of potential interest for aquaculture. The total lipid contents (TL) of muscle and liver of the captive fish were 2.5-fold greater than those of the wild fish. In consequence, contents of triacylglycerols were much higher in tissues of the captive fish. Distribution of fatty acids in total lipids and lipid classes of muscle and liver was also different between both groups of fish. For instance, percentages of 20:4n−6, 20:5n−6 and 22:6n−3 were considerably higher in the wild fish, whereas 18:1, 20:1, and 22:1n−9 as well as 18:2n−6 and 20:5n−3 were more abundant in the captive fish. These results suggest that the lipid composition of the commercial diet supplied to the captive black seabream differed greatly from that of the diet consumed by the fish in the wild, which hypothetically contains the desirable composition for the lipid nutrition of this fish species. Despite the good growth and survival achieved by the black seabream after one year in captivity, the significant accumulation of lipids and the imbalance of essential fatty acids in their muscle and livers, together with the absence of spawning, suggest that future research on the lipid requirements of this omnivorous species is necessary in order to establish whether the administration of currently available aquaculture formulated feeds is adequate for good black seabream performance and reproduction.     

Welfare status of cultured seabass (Dicentrarchus labrax L.) and seabream (Sparus aurata L.) assessed by blood parameters and tissue characteristics

Seabass (SBS) and seabream (SBM) juveniles were reared with the goal of obtaining three different final densities (SINT = 40 kg m^?3; INT = 20 kg m^?3; SEM = 0.2 kg m^?3) to ascertain the effects thereof on the welfare of the fish. Significant blood metabolites and hepatic glycogen were determined every 3 months and at harvest. Trials lasted 18 months in seabass and 17 months in seabream. At the end of experiment the main biometric productive parameters and quality of body composition were also recorded. Regarding intensively reared seabass (SINT‐SBS, INT‐SBS), the plasma triglycerids, total cholesterol and transaminases (AST, ALT) were always significantly higher than in semi‐intensively maintained fish (SEM‐SBS). At the final sampling in the SINT‐SBS batch, the total protein and glucose were also markedly increased. Conversely, at harvest the liver glycogen content decreased in SINT‐SBS (34 ± 8 mg g^?1 liver) with respect to INT‐SBS (57 ± 12 mg g^?1 liver) and SEM‐SBS (63 ± 11 mg g^?1 liver). No differences among groups were observed for creatine kinase (CK) and lactate dehydrogenase (LDH). With regard to seabream, SINT‐SBM and INT‐SBM constantly showed plasma triglycerids and total cholesterol as being significantly higher when compared to SEM‐SBM. In the final two blood samplings, SINT‐SBM exhibited the most elevated values for LDH. At harvest, AST, ALT, total protein and glucose markedly increased in SINT‐SBM, whereas liver glycogen content was reduced (22.5 ± 9 mg g^?1 liver), more than in INT‐SBM (70 ± 16 mg g^?1 liver) and SEM‐SBM (75 ± 20 mg g^?1 liver). In both seabass and seabream, body composition was very similar in the different stocking densities, except for total cholesterol. Total n‐3 polyunsaturated fatty acid (PUFA) in seabream was significantly different from fish of the semi‐intensive groups; however, nutritional values and fatty acid profiles were equally good. The intermediate final density of seabass and seabream at 20 kg m^?3 seemed to give the best results in terms of their well being when compared to fish reared at 40 kg m^?3. The absence of differences in blood metabolites and hepatic glycogen levels between the intensive batch and the semi‐intensive groups until harvest was a reference to the positive status of the fish. A density of 20 kg m^?3 can be considered acceptable for farm strategy planning for raising healthy on‐growing seabass and seabream juveniles.     

Reared perch v. wild perch – first results about organoleptical and technological quality

The Eurasian perch, Perca fluviatilis L., has good potential as a candidate for inland aquaculture diversification in Europe. To aid the development of this new product, we define a quality reference based on attributes of wild perch, and compare the quality of perch reared in three different culture systems: (1) extensive (pond), (2) semi‐extensive (tank) and (3) intensive (re‐circulation system). In order to define a precise quality reference, it is necessary to characterize the spatial and seasonal variability in quality attributes of wild fish. To do so, wild perch were harvested every three month for a full biological cycle from two different natural systems (Lake Geneva in France and the River Rhine in the Netherlands). Two components of quality were taken into account: organoleptical (colour of skin and fillets (L*C*H° system; 12 variables), seven morphological measurements) and six technological components (viscero, hepato and gonadosomatic index, perivisceral fat index, fillet yield and condition factor K2). Using Principal Component Analysis, we show that within‐season variability is high, particularly for Rhine perch. Perch from the Rhine have a compact body and caudal fin, and possess a longer mouth and head than perch from Lake Genva. Reared fish possess wild‐like characteristics except for technological variables. Fish from extensive and semi‐extensive system show similar attributes, while fish from intensive rearing systems show a higher viscerosomatic and perivisceral fat index.     

Effects of hatchery rearing on Florida largemouth bass Micropterus floridanus resource allocation and performance under semi‐natural conditions

This study examined the growth, activity, metabolism and post‐release survival of three groups of Florida largemouth bass Micropterus floridanus: wild‐caught fish, hatchery fish reared according to standard practice (hatchery standard) and hatchery fish reared under reduced and unpredictable food provisioning (hatchery manipulated). Hatchery‐standard fish differed from wild‐caught fish in all measured variables, including survival in semi‐natural ponds. Hatchery‐standard and hatchery‐manipulated fish showed higher activity levels, faster growth and lower standard metabolic rates than wild‐caught fish in the hatchery. Fish reared under the manipulated feeding regime showed increased metabolic rates and increased post‐release growth, similar to wild‐caught fish. Their activity levels and post‐release survival, however, remained similar to those of hatchery‐standard fish. Activity was negatively correlated with post‐release survival and failure of the feed manipulation to reduce activity may have contributed to its failure to improve post‐release survival. Activity and post‐release survival may be influenced by characteristics of the rearing environment other than the feeding regime, such as stock density or water flow rates.     

Morphological differences between wild and farmed Mediterranean fish

Gilthead seabream (Sparus aurata L.) and European seabass (Dicentrarchus labrax L.) are important commercial marine fish species both for aquaculture and fisheries in the Mediterranean. It is known that farmed individuals escape from farm facilities, but the extent of escape events is not easy to report and estimate because of the difficulty to distinguish between wild and farmed individuals. In this study, significant differences provided through morphometry evidence that the cranial and body regions of seabream and seabass are different regarding their farm or wild origin at different scales. Morphological variations have been shown to be a valuable tool for describing changes in shape features. Therefore, the biomass contribution of escapees to local habitats could be determined by identifying escaped individuals from fisheries landings as a first step to assess the potential negative effects of fish farm escapees on the environment, and their influence on wild stocks and local fisheries.     

Impact of live food on survival and growth of hatchery‐reared sea trout (Salmo trutta trutta L.) parr in the wild

Survival rates and growth parameters of hatchery‐reared sea trout (Salmo trutta trutta L.) fry were determined after stocking in the wild. The larvae were hatchery‐reared for 12 weeks in two groups: fry were fed either on live zooplankton and live chironomidae larvae (LFG), or fed a pellet diet (PFG). The survival rate and specific growth rates were higher in the LFG than in the PFG group. Most effective for hatchery‐reared fish intended for stocking was the natural, live feed. The mean number of chironomid larvae found in the stomachs of fish that were initially captured in the wild was significantly higher in the LFG than in the PFG group. The live diet supplied in the rearing period had a positive impact on the foraging skills of the sea trout fry and their survival in the wild after their release on 24 April 2010.     

Rearing environment influences boldness and prey acquisition behavior,and brain and lens development of bull trout

Animals reared in barren captive environments exhibit different developmental trajectories and behaviors than wild counterparts. Hence, the captive phenotypes may influence the success of reintroduction and recovery programs for threatened and endangered species. We collected wild bull trout embryos from the Metolius River Basin, Oregon and reared them in differing environments to better understand how captivity affects the bull trout Salvelinus confluentus phenotype. We compared the boldness and prey acquisition behaviors and development of the brain and eye lens of bull trout reared in conventional barren and more structurally complex captive environments with that of wild fish. Wild fish and captive reared fish from complex habitats exhibited a greater level of boldness and prey acquisition ability, than fish reared in conventional captive environments. In addition, the eye lens of conventionally reared bull trout was larger than complex reared captive fish or same age wild fish. Interestingly, we detected wild fish had a smaller relative cerebellum than either captive reared treatment. Our results suggest that rearing fish in more complex captive environments can create a more wild-like phenotype than conventional rearing practices. A better understanding of the effects of captivity on the development and behavior of bull trout can inform rearing and reintroduction programs though prediction of the performance of released individuals.     

The role of social learning in conservation and fisheries reintroductions

The plight of the world fish stocks is all too well documented. As part of an ongoing attempt to bolster fish stocks for both commercial and conservation purposes, many fish are reared in captivity and released into the wild. It is well known that hatchery‐reared fish have low post‐release survival compared with wild fish of similar age. Part of the reason for this high mortality is that hatchery fish show deficits in virtually all aspects of their behaviour, including prey selection and predator avoidance. Much behaviour requires repeated experience so that it may become fine‐tuned to prevailing circumstances via learning during development. It has been suggested that inappropriate behaviour is encouraged when fish are reared in the unnatural surroundings of the hatchery. However, hatchery fish can be taught to recognise live, novel prey items and predators and the rate of learning is increased in the presence of a more knowledgeable conspecifics. Here we present data showing how social learning protocols can be used to dramatically increase foraging success in juvenile Atlantic salmon. We also outline related aspects of our ongoing research and discuss some of the possibilities for altering hatchery practices to maximize post‐release survival.     

The histological analysis,colorimetric evaluation,and chemical quantification of melanin content in ‘suntanned’ fish

Human melanocytes respond to UV irradiation by increasing the synthesis of melanin. While much is now understood of the pathways governing this process and the nature of the melanin synthesized, little is known of melanins produced by lower vertebrates and their capacity to respond to UV. Here we report that a fish, red seabream, can undergo ‘suntanning’. Histological, colorimetric and chemical assays were performed for suntanned red seabream fish bred in net cages to analyse the melanins and compared with shaded or wild red seabream fish. For color evaluation, the L* values of suntanned fish were dramatically lower than those in the other two groups. Pyrrole‐2,3,5‐tricarboxylic acid (PTCA), an indicator of eumelanin, was detected in suntanned fish at five times higher levels than in shaded or wild fish while 4‐amino‐3‐hydroxyphenyl‐alanine (4‐AHP), a marker for pheomelanin, could not be detected in any of the samples. Histological analysis showed that melanocytes in the suntanned skin enlarged and increased in number to form a monolayer at the surface of the skin. Analysis of L* values and PTCA levels showed quite a high correlation coefficient (r = ?0.843). When comparing shaded and wild red seabream fish, the scores were closer but some significant differences were still found in some body areas. These results indicate that eumelanin accumulates in suntanned fish during the increase in skin color, which is induced by sunlight, presumably by ultraviolet radiation.     

Environmental rearing conditions produce forebrain differences in wild Chinook salmon Oncorhynchus tshawytscha

Recent studies suggest that hatchery-reared fish can have smaller brain-to-body size ratios than wild fish. It is unclear, however, whether these differences are due to artificial selection or instead reflect differences in rearing environment during development. Here we explore how rearing conditions influence the development of two forebrain structures, the olfactory bulb and the telencephalon, in juvenile Chinook salmon (Oncorhynchus tshawytscha) spawned from wild-caught adults. First, we compared the sizes of the olfactory bulb and telencephalon between salmon reared in a wild stream vs. a conventional hatchery. We next compared the sizes of forebrain structures between fish reared in an enriched NATURES hatchery and fish reared in a conventional hatchery. All fish were size-matched and from the same genetic cohort. We found that olfactory bulb and telencephalon volumes relative to body size were significantly larger in wild fish compared to hatchery-reared fish. However, we found no differences between fish reared in enriched and conventional hatchery treatments. Our results suggest that significant differences in the volume of the olfactory bulb and telencephalon between hatchery and wild-reared fish can occur within a single generation.     

Effects of Hatchery Rearing on Brain Structures of Rainbow Trout, Oncorhynchus mykiss

In this study, we contrast brain morphology from hatchery and wild reared stocks to examine the hypothesis that in salmonid fishes, captive rearing produces changes in brain development. Using rainbow trout, Oncorhynchus mykiss, as a model, we measured eight regions of the salmonid brain to examine differences between wild and hatchery reared fish. We find using multiple analysis of covariance (MANCOVA), analysis of covariance (ANCOVA) and discriminant function analysis (DFA) that the brains of hatchery reared fish are relatively smaller in several critical measures than their wild counterparts. Our work may suggest a mechanistic basis for the observed vulnerability of hatchery fish to predation and their general low survival upon release into the wild. Our results are the first to highlight the effects of hatchery rearing on changes in brain development inbreak fishes.