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Widely appreciated in gastronomy, many harvested and farmed Crustacean species are a great economic resource in the fishery business, throughout the world. Factors like pollutants and/or improper management in on-board/on-shore practices may adversely affect their edible quality and shelf-life. Recent studies from applied scientific research have been supporting the fishing industry and trading companies, in order to improve post-harvest practices and offer a safe and competitive product of high quality. Here, we developed an innovative molecular approach to get basic information on crustacean quality by using the European lobster Nephrops norvegicus (Linnaeus, 1758). For the first time, we investigated muscle-exudates of thawed lobsters for the presence of nucleic acids and analysed mRNA targets selected at NCBI to test their potential use as biomarkers of quality in frozen-thawed seafood. By RT-PCRs and sequencing we validated small amplicons of Actin, Cytochrome oxidase subunit 1 (COI), Calpain M, Prophenoloxidase (ppo) and Crustin-like antimicrobial peptide. In comparative One Step RT-PCR and/or One Step RT-qPCR assays, we analysed their amplification profiles at 1, 4 and 7 days after thawing (a.t.), in lobsters maintained to checked temperatures from capture. Based on amplification efficiency (AE), inversely correlated to the time after thawing and loss of RNA targets integrity, we identified new highly sensitive, species specific early indicators of quality. Our results highlight the advantage of using amplicons from exudates in different food science applications including authenticity/traceability, and provide new insights for the development of scientific tools applicable to crustacean quality monitoring on industrial-scale.

Management of dredged materials disposal is regulated by several environmental normative requirements, and it is often supported by the integration of chemical data with ecotoxicological characterization. The reliability of a bioassay to assess the potential toxicity of dredged sediments requires the selection of quality criteria that should be based on simple analytical methods and easily understandable hazard for politicians and environmental managers. The sea urchin embryo-toxicity bioassay is considered an essential component for evaluating the quality of sediments in harbour areas but its use, when based exclusively on the observation of normal vs. abnormal embryos, may alter the interpretation of the results, overestimating the risk assessment. To improve the reliability of this assay in establishing a causative relationship between quality of sediments and sea urchin embryonic development, here we developed and validated three Integrative Toxicity Indexes (ITI 2.0, ITI 3.0, ITI 4.0), modifying the already-known ITI (here ITI 1.0). Based on this aim, we used Taranto harbour as a model pilot-study to compare results to those obtained from standard criteria. Among the tested indexes, the ITI 4.0, discriminating strictly developmental delay and morphological defects from fertilized egg to gastrula stage, resulted in the most promising.

The sea urchin embryo develops a well-defined biomineralized endoskeleton, synthesized exclusively by the skeletogenic cells, supported by ectodermal cues for the correct skeleton patterning. The biomineralization process is tightly regulated via a hierarchical order of gene expression, including transcription and growth factors, biomineralization proteins. Recently, the role of kinases and intracellular signaling pathways in sea urchin skeletogenesis has been addressed, although the downstream components still remain unknown. In this study, we investigated the role of phosphatidylinositide 3-kinase (PI3K)-mediated signaling pathway in Paracentrotus lividus, to identify its genes/proteins targets. The effects of LY294002 (LY), a PI3K-specific inhibitor, were evaluated at morphological and molecular levels. Treatment with 40 μM LY from the blastula stage completely blocked skeleton deposition, which was reversed by wash out experiments. Besides, LY caused a slight delay in the tripartite gut development. Despite the skeleton absence, a few skeleton-specific proteins/mRNAs were regularly expressed and localized in LY-treated embryos, as shown for MSP130 and SM50 by immunofluorescence and in situ hybridization experiments. QPCR analyses showed that LY differently affected the expression of genes coding for other biomineralization proteins, transcription and growth factors. SM30 and carbonic anhydrase expression was severely downregulated, while almost all the transcription factors analyzed were upregulated. Based on the present results and in silico analyses, we propose an “interactomic” model simulating PI3K connections in P. lividus embryos. Our findings define a novel regulatory step in the embryonic skeletogenesis, and provide valuable molecular data for further studies on the role of PI3K signaling in invertebrate biomineralization.

Submersible sea-cages represent an alternative method to standard surface-based cages, and could help solve several production problems that exist in surface-based farming, such as heavy storms, algal and jellyfish blooms and attacks by predators. In this study, we investigated growth, haematological, biochemical and immunological parameters of European sea bass ( Dicentrarchus labrax ) farmed in surface and submersible cages, to compare the physiological status under different rearing conditions. In addition, we evaluated the use of erythrocyte HSP70 levels as a biomarker of fish welfare. The study was conducted in two submerged and two surface cages filled with D. labrax . No significant differences were observed in the growth rate and the relative condition factor between the two groups. Significantly higher levels of serum cortisol, haematocrit plasma glucose and total plasma proteins were measured in surface-based cages. Conversely, fish in submerged cages showed higher haemolytic activity and lysozyme levels in both mucus and kidney than surface reared sea bass. Results obtained in this study indicate that submergence could be a favorable condition for sea bass farming, suggesting mariculture in submerged net cages as a promising system that allows producers to minimize stress (included that from predation by fish-eating birds) and therefore favor animal welfare.