Amplification efficiency of RNA sequences related to stress and immune response gene as an innovative approach in the quality evaluation of the crustacean Nephrops Norvegicus (Linnaeus, 1758)

Amplification efficiency of RNA sequences related to stress and immune response as an innovative approach in the quality evaluation of the crustacean Nephrops norvegicus (Linnaeus, 1758) C. Costa, A. Pinsino, R. Bonaventura, R. Russo, F. Zito, V. Matranga Istituto di Biomedicina e Immunologia Molecolare "A. Monroy", CNR, Palermo, Italy The lobster Nephrops norvegicus has a relevant economic value in the crustacean business sector. Environmental pollution or improper management of the cold chain may adversely affect quality and shelf life of crustaceans. World fishing industry, seafood markets and consumers are strongly requiring new scientific tools for the assessment of lobster quality and competitiveness. In this work we developed an innovative molecular approach to assess the shelf life of the lobster N. norvegicus. For the first time we investigated muscle exudates of thawed specimens as potential source of nucleic acids. Despite their hypothesized degradation, we were able to detect specific nucleotide sequences. By a search at NCBI database, we selected as potential indicator(s) of quality, RNA sequences validated in other species for their responsiveness to pollutants. By RT-PCR, we amplified small amplicons of the following sequences coding for stress and immune response genes: Actin, Cytochrome Oxidase I (COI), Calpain M, Prophenoloxidase (ppo) and Crustin-like antimicrobial peptide. By comparative One Step RT-PCR and/or QRT-PCR assays we evaluated the amplification efficiency (AE) of RNAs from thawed specimens, maintained at 4°C for 0, 1, 2, 3 and 7 days (shelf-life). We found that long shelf-life and lowest quality correlate with the lowest values of AE. Our results provide an opportunity for food science applications on industrial-scale management to monitor over time quality and shelf-life. Financed by the DeCroMed Project, EU - Research & Innovation Strategy - Sicily Region - PO-FESR 2007- 2013, CUP: G93F12000190004. We acknowledge the technical assistance of Mr. M. Biondo.