The pollutant load was monitored in an intensive on-shore plant; evaluation especially concerned one basin, during a period of time in which the water temperature (21-23°C), the stocking density (25 kg/m3), the loading and unloading flow (40 dm3/s) remained steady at the highest values. The results show that the maximum pollution levels are characterized by high concentrations of suspended organic materials, as well as by high BOD5/COD rates; moreover, more than 60% of the suspended solid waste consists of particles with a maximum dimension ranging from 200 μm to more than 500 μm. Tests show that an effluent channel is less efficient in reducing the pollutant load of waste water than sedimentation treatment. The laboratory sedimentation trials showed - for a period of about 3 hours - minimum and maximum solid particle sedimentation speeds of 2.56 mm/s and 46.8 mm/s respectively, which should be considered when deciding the turbid speed and the dimensions of basins for this type of effluent. In any case, the sedimentation process carried out in the laboratory was only just sufficient to bring the farm effluent within the legal limits. Therefore mechanical filtration treatment may suit the specific requirements of the farm waste water used for the experimental trials. A 50 μm mesh would be considered sufficient for any critical situation.

Currently available data show that shellfish and finfish production in Italy, derived both from fisheries and aquaculture activities, is on the order of 474,000 tons, each activity representing 50 % of the total amount. In this context, the finfish aquaculture industry contributes on average 31 % to the national aquaculture production and on average 59 % of its value, giving a total amount of 72,000 tons and a value of around 351 million € (2010). According to FEAP statistics, Italy is the fourth largest finfish producer in EU27, after the UK, Greece, and Spain, while it is also one of the six largest finfish producers among the non-EU and EU member countries, together with Norway, UK, Greece, Turkey, and Spain. Presently, fish culture activities are mainly focused on rainbow trout (Oncorhynchus mykiss, 55.5 %), followed by European sea bass (Dicentrarchus labrax, 13.6 %), gilthead sea bream (Sparus aurata, 12.2 %), gray mullet (Mugil cephalus, 5.3 %), sturgeon (Acipenser spp., 2 %), and European eel (Anguilla anguilla, 1.7 %). Over the last 20 years, freshwater fish production and aquaculture (trout, carp, and eel) have decreased in Italy, with the exception of sturgeon. In contrast, marine fish production has significantly increased during the same period, and the two leading species, European sea bass and gilthead sea bream, presently contribute 25.8 % of the finfish production. From 1,900 tons in 1990, production reached 19,000 tons in 2010, with a 900 % increase, at an average percentage of 4.5 %. In addition, new marine fish species were successfully cultured over the same period. This review outlines the past and present situation of finfish culture in Italy and discusses future developments and priorities, with particular emphasis on new, emerging aquaculture species.

Diablo (or SMAC) is a protein released from mitochondria following apoptotic stimuli and inhibits the actions of Inhibitors of Apoptosis (IAP) proteins. IAPs regulate the activity of caspases and NFkB, the primary executioners of apoptosis and of inflammation, respectively. Thus, Diablo is important for the regulation of cellular responses to damage. In Northern Europe, statutory governmental marine monitoring programs measure various biomarkers in flounder to indicate biological effects of pollutant exposure. More recently transcriptomic techniques have been applied in flounder to gain a more comprehensive understanding of pollutant effects, and to discover novel biomarkers. In most of these studies utilising flounder, Diablo was amongst the most highly increased transcripts identified. The aim of this study was to further examine piscine Diablo, at the gene level and mRNA level, after exposure to prototypical pollutants, and in flounder caught from polluted environments. The results show that two genes encoding Diablo exist in fish species, and in flounder one of these genes is increased in liver after exposure to polyaromatic hydrocarbons and polychlorinated biphenyls, and also in livers from fish living on contaminated estuarine sediments. Therefore, Diablo measurement has potential as a biomarker of pollutant exposure, and could indicate damaging effects of chemical contaminants. (C) 2012 Elsevier Inc. All rights reserved.

Aflatoxins are one of the most widespread and worrisome sources of feed contamination worldwide, and have a considerable impact on fish farm production, leading to high mortality and a gradual decline in fish stock quality in aquaculture. In this study, we investigated, for the first time, the effects induced in vitro by aflatoxin B-1 (AFB(1)) on Sparus aurata hepatocyte culture and we compared our results with the Microtox (R) system using Vibrio fischeri. At AFB(1) doses ranging from 1 to 10 mu g.mL(-1), the results showed signs of primary necrotic cell death in hepatocytes and a very toxic evaluation with Microtox (R); between 0.005 and 1 mu g.mL(-1), the cytotoxic effects and apoptotic delayed death in eukaryotic cells corresponded with an evaluation of no toxicity or biostimulatory effect using V. fischeri. Overall, our results highlighted equivalent toxic responses and overlapped with values of EC50/IC50. Hence, these two in vitro systems could be considered as a useful starting point in the design of new batteries to evaluate the toxicity of potentially dangerous feed-borne substances.

Among all know mycotoxins, aflatoxin B1 is one of the most studied for its hepatotoxic, carcinogenic, mutagenic, teratogenic, and immunosuppressant effects. However, metabolic and toxicological studies on aflatoxins in farmed Sparus aurata are limited and restricted to in vivo trials. This work aimed to study the effects of AFB1 acute and chronic exposure on CYP1A and GST enzymes induced in vitro on S. aurata hepatocytes by immunoblot analysis, thus relating the cytotoxic effects leading to cell death by apoptotic studies. Immunofluorescence analysis revealed that cell damage was not recoverable but permanent, as the cellular repair systems were unable to recover the induced toxic insult. Our results showed detection of several CYP1A bands, enlightening an indirect correlation between induction of CYP1A with dose and time of exposure. The decreased expression of CYP1A over prolonged exposure times, along with high toxic concentration, could be related with lethal damage observed on hepatocytes by contrast phase and immunofluorescence analysis. A particular pattern of expression was found for GST isoforms upon AFB1 exposure, identifying each isoform profile two different kind of toxic insult. The 65 KDa and the 49 KDa bands being suggestive for markers of acute and chronic response respectively. Interestingly, apoptosis induction, considered an early lesion to DNA, was found associated with the chronic damage along with the low toxic concentration. The new cell model from S. aurata has been proven to be a useful and valid tool to further investigate the modulated response of liver phase I and II enzymes to AFB1.

In the present study, cytotoxic effects of the polycyclic aromatic hydrocarbons benzo[a] pyrene (B[a]P) were investigated in Sparus aurata hepatocytes primary cultures after acute and chronic exposure. Cells were treated with a wide range of B[a]P doses (1 pg/mL to 100 µg/mL) for 24, 48 and 72 h. B[a]P toxicity was quantified in sea bream hepatocytes by MTT assay and immunofluorescence analysis of apoptosis after the various exposure periods, in order to evaluate the hepatic damage and toxicity range. Results showed three cytotoxic responses: B[a]P cell death for primary necrosis after exposure to high concentrations for short times, apoptosis induction with the use of sublethal doses and cell proliferation allied with neoplastic foci formation after exposure to low concentrations for long times. This responses provided an interesting correlation between the damage caused on hepatocytes and the metabolism of this toxic compound, to date mainly studied in vivo. Additionally, the statistical analysis revealed that the effects of time and dose were significant for both parameters and especially the time was extremely significant (P<0.0001), in fact B[a]P induced damage that increased over time. Our findings demonstrated and confirmed that S. aurata is a very sensitive species to B[a]P exposure since adverse effects were found at all tested doses. Furthermore, the new in vitro animal model can be considered a useful tool for studying the cellular effects induced by any contaminant harmful for farmed fish.

The main goal of this work was to determine the effect of dietary live yeast Saccharomyces cerevisiae on the oxidative status of sea bass Dicentrarchus labrax juveniles. Fishes were fed on three diets: the GM group were fed a diet containing lyophilized yeast grown on grape must, the CS group were fed a diet containing lyophilized yeast grown on cornstarch, and the control group were fed a diet without yeast. The activity of the main antioxidative enzymes, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase, glutathione S-transferase (GST), and glutathione (GSH) content, as well as lipid peroxidation, was measured in the liver of sea bass juveniles 90 days after hatching. Supplementation of the diet with S. cerevisiae significantly reduced the SOD and CAT activity, increased the GST activity, decreased the GSH content, and had no effect on lipid peroxidation. The results support the already reported radical-scavenging properties of yeast and usefulness of its employment as antiperoxidative agent in fish.

The effects of lycopene-enriched extenders on in vitro quality characteristics and lipid peroxidation of turkey semen after both chilled and frozen storage were evaluated. Five pools of semen diluted in extenders containing 0, 0.05 or 0.1 mg/mL lycopene were stored either at 5°C for 48 h or cryopreserved as pellets. Mobility, viability, osmotic resistance, DNA integrity as well as lipid peroxidation (malonaldehyde production) of spermatozoa were evaluated in fresh, chilled and frozen sperm. Semen quality was generally reduced after storage, especially post-freezing. However, in semen extended with the highest dose of lycopene, neither viability and osmotic-resistance of chilled sperm, nor DNA integrity of frozen sperm, differed significantly from those of fresh semen. Lipid peroxidation was higher in refrigerated than in fresh or cryopreserved spermatozoa. Anyway, spermatozoa chilled in lycopene-enriched extenders had significant lower malonaldehyde levels than those chilled without lycopene, whereas in frozen semen the addition of lycopene contributed to maintain the lipid peroxidation no different from that scored in fresh semen. In conclusion, lycopene improved the survival of turkey spermatozoa after liquid-storage and protected their DNA integrity against cryodamage. The positive effect of lycopene addition to extenders was likely due to its role in limiting the amount of sperm lipid peroxidation after both refrigeration and cryopreservation.

The development of primary cultures and cell lines from aquatic organisms is a valuable tool for a wide range of research activities applied to aquaculture. Despite several efforts, derivation and long-term culturing of primary hepatocytes from marine vertebrates are still rare and unsuccessful. This is the first report to fully characterize long-term cultures of primary hepatocytes from the European seabream, Sparus aurata L. (Osteichthyes, Sparidae) (SaHePs). In this new model, hepatocyte cells were long-term viable, active proliferating, and fully retained liver function up to 3 weeks. SaHePs expressed a differentiated phenotype, owing to the reacquisition of the peculiar cytoarchitecture with the complete assembly of cytoskeletal and junctional network, as shown by the production and immunolocalization of several polarity markers and cytoskeletal proteins (MDR1, ZO-2, C-CAM1, Vimentin, Cadherin, beta-Tubulin, beta-Catenin, beta-Actin). Cytostructural analysis to identify polarized expression and bile canaliculi formation was performed by immunofluorescence and contrast phase microscopy. Long cultured SaHePs also demonstrated evidence of Albumin, alpha 1-Antitrypsin (AAT) and alpha-Fetoprotein (AFP) synthesis, expression of the detoxifying metabolic enzyme cytochrome P-4501A (CYP 1A), and production of hepatocyte specific cytoskeleton proteins, such as Cytokeratin 8 (CK8) and Cytokeratin 18 (CK 18). The presence of specific markers for hepatic phenotype, detected by immunocytochemistry and Western blot analysis, is suggestive of the full maintenance of a highly differentiated phenotype and hepatic maturation. These data demonstrate that SaHePs can be long cultured without losing the hepatic functionality. This study provides a useful tool for innovative research applications in fish toxicological, pathological, and physiological studies, as one of the few hepatic, functionally active, in vitro model from marine fish.

Contractions of ovarian tunica albuginea, the teleostean cystovary wall layer containing smooth muscle fibres, facilitate oocytes and fluids movements within the ovary, oocytes ovulation and spawning. Fish isotocin, the homologue hormone of mammalian oxytocin, plays a significant role in ovulation, oviduct contraction and spawning. In the present study, ovarian wall spontaneous contraction, as well as isotocin in vitro effect on tunica albuginea contractility, was analysed in female seabream in different reproductive conditions: vitellogenesis, regressing (post-spawning) and extensive atresia. Tunica albuginea spontaneous contractility was recorded using ovary wall strips mounted in an organ bath containing modified Ringer's solution. The strips were then exposed to cumulative doses of isotocin (6, 30, 60 μg/ml). Female seabream in regressing condition exhibited the highest level of tunica albuginea spontaneous contraction amplitude compared with the other two groups. Only fish in vitellogenesis state showed a significant increase in contraction amplitude after isotocin administration at the dose of 30 μg/ml. The same group exhibited also a significant isotocin dose-dependent decrease in the contractile frequency. These results confirm the involvement of isotocin in stimulating tunica albuginea contractile activity during the oestrogen-regulated phase of vitellogenesis, whereas the absence of significant effects of isotocin on ovarian contractility in fish at the regressing state might be ascribed to the occurrence of a contractile activity autonomously regulated by the internal pacemaker system. The absence of exposed isotocin receptors could explain the lack of effects of the isotocin administration in seabream showed extensive atresia of the follicular cells.

The effects of lycopene-enriched extenders on motility, viability, osmotic resistance, DNA integrity and lipid peroxidation of rabbit sperm were examined after both chilled and frozen storage. Five pools of semen diluted in extenders containing 0, 0.05 or 0.1. mg/mL of lycopene were refrigerated at 5°C for 48. h or cryopreserved. Sperm quality was generally compromised after storage, especially post-freezing, however lycopene limited the amount of sperm lipid peroxidation after chilling and freezing. In chilled sperm, the highest dose of lycopene was provided to maintain the viability, acrosome and DNA intactness similar to that of fresh semen and contained the reduction of sperm motility, whereas in cryopreserved semen lycopene protected the DNA integrity of sperm, even if not in a dose-dependent way. So lycopene diminished sperm lipid peroxidation during refrigeration and cryopreservation, prolonging the survival of rabbit sperm after liquid storage, but it had a limited effect on sperm cryosurvival. © 2012 Elsevier B.V.

The purpose of the present study was to evaluate the relationship between the feeding frequency and timetable on egg parameters and reproductive performance of broiler breeder hens. The study involved 480 Ross-308 strain broiler breeder hens from 31 to 38 weeks of age. Birds were randomly assigned to 15 dietary treatments, including four replicates (seven hens and one rooster/treatment), and fed conventional diet (11.7 MJ kg-1 of metabolisable energy, 14% crude protein). Treatments were as follows: (1) once-a-day feeding schedule (control), in which birds received their total allocated feed at 0400 h; (2) twice-a-day feeding schedule, in which the daily feed allowance was divided into two equal meals (50:50) and fed at 0400 and 1600 h; (3) twice-a-day feeding, 75:25 at 0400 and 1600 h; (4) twice-a-day feeding, 50:50 at 0400 and 1200 h; (5) twice-a-day feeding, 75:25 at 0400 and 1200 h; (6) twice-a-day feeding, 75:25 at 0600 and 1600 h; (7) twice-a-day feeding, 50:50 at 0600 and 1600 h; (8) three times feeding schedule, 50:25:25 at 0400, 1200 and 1600 h; (9) three times feeding schedule, 33:33:33 at 0400, 1200 and 1600 h; (10) three times feeding schedule, 25:50:25 at 0400, 1200 and 1600 h; (11) three times feeding schedule, 25:25:50 at 0400, 1200 and 1600 h; (12) three times feeding schedule, 50:25:25 at 0600, 1200 and 1600 h; (13) three times feeding schedule, 33:33:33 at 0600, 1200 and 1600 h; (14) four times feeding schedule, 33:22:22:22 at 0400, 0800, 1200 and 1600 h; and (15) four times feeding schedule, 50:17:17:17 at 0400, 0800, 1200 and 1600 h, respectively. According to our results, until 38 weeks of age the egg production, egg-shell weight, albumen height and yolk colour as well as Haugh unit were positively influenced (P < 0.05) in the hens fed twice a day. Similarly, also using the evaluation index and sub-ordinate function methods, the higher scores were obtained by birds fed more than once a day. In contrast, all feeding regimens did not affect the other egg parameters. Therefore, it can be concluded that feeding broiler breeder hens twice or three times per day improved the egg traits and reproductive performance during the egg peak production phase.

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Shellfish aquaculture is a widespread activity in the Italian peninsula. However, only two bivalve species are mainly cultured along the coastline of that country: the Mediterranean mussel Mytilus galloprovincialis and the Manila clam Venerupis philippinarum (Ruditapes philippinarum). By contrast, just a few other mollusc species of commercial interest are scarcely reared at a small-scale level. After analysing the current status of Italian shellfish production, this paper reports and discusses the potential for culturing several different invertebrate species [i.e., the European flat oyster Ostrea edulis, the grooved carpet shell Venerupis decussata (Ruditapes decussatus), the razor clams Ensis minor and Solen marginatus, the cephalopod Octopus vulgaris, and the purple sea urchin Paracentrotus lividus] in this country. In addition, a detailed overview of the progress made in aquacultural techniques for these species in the Mediterranean basin is presented, highlighting the most relevant bottlenecks and the way forward to shift from the experimental to the aquaculture phase. Finally, an outlook of the main economic and environmental benefits arising from these shellfish culture practices is also given.

Neuroglia has been historically considered the “glue” of the nervous system, as the ancient Greek name suggests, being simply referred as non-neuronal cells, with supporting functions for neurons in the CNS of mammalian and lower vertebrates. All around the world, approximately 283 cell lines were obtained from fish, yet none of these was from the brain of Sparus aurata, neither in cell lines nor as primary culture. Here we describe a novel in vitro reproducible neuroglial marine model for establishing primary neuroglial cell cultures, by dissociating the whole brain of seabream juveniles. We showed that proliferating neural stem cells produced alongside three generating lineages, such as neuronal precursor cells, astroglial precursor cells and oligodendroglia precursor cells, which developed respectively neurons, astrocytes and oligodendrocytes. The radial glia, finely described by morphological studies and immunochemical antigen expression, showed a peculiar spatial distribution, giving rise simultaneously both to astrocytes and neuronal precursors within a highly proliferative assemblate. Radial glia cells were assessed by glial fibrillary acidic protein (GFAP) and vimentin reactivity, astrocytes by GFAP, neurons by the neuron-specific markers for ubiquitin carboxy-terminal hydrolase 1 (UCHL1) and intermediate filament associated protein (NF), whereas myelinating oligodendrocytes were immunostained with anti-myelin basic protein (MBP) and anti-O4. Our findings suggest that seabream neuroglial cells gain in 3-4 weeks of culturing proliferation, neuroglial differentiation, and oligodendrocyte maturation with myelination, thus disclosing on the possibility that mixed neuroglial cultures can accelerate the maturation of oligodendrocytes and the regeneration of CNS injury in fish.

Over the past few years, consumers have been increasing their awareness about environmental, health and safety concerns, gradually changing their habits in favour of organic food. In Europe, the organic aquaculture is legislated by Comm ission Regulation (EC) 710/2009. In Italy, only ten farms are involved in the supply of certified organic fish and only some pilot projects were carried out for organic farming of different species. Therefore, the purpose of this research was to evaluate o rganic technical feasibility and production costs, comparing them with conventional production. This study contains the first considerations about organic production and its relative costs for seabass fry, one of the most reared species of the country. Con version to organic production naturally involves additional costs concerning the conversion process itself, the production of a new type of product and the lower output. In fact, results have shown that certification and feed costs represented the most sig nificant difference between conventional and organic production. .

La società FIND si occupa di sostenere lo sviluppo e l'innovazione nel settore agroalimentare, ed in particolare nell'ambito dell'acquacoltura e della pesca, attraverso l'ottima gestione e l'utilizzo di tecnologia sempre all'avanguardia, fornendo tutta l'esperienza del nostro team tecnico e scientifico. La società offre una consulenza professionale e lo sviluppo di progetti personalizzati nel settore di riferimento, grazie alla presenza di un personale versatile e degli stessi partner che fanno parte della società. Il nostro lavoro è indirizzato sia agli istituti pubblici che alle aziende private, alle organizzazioni governative e non, ai centri di ricerca nazionali e internazionali.