A new molecular approach for shelf-life evaluation of the crustacean Nephrops norvegicus (Linnaeus, 1758)

A new molecular approach for shelf-life 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 European lobster Nephrops norvegicus is an appreciated seafood with a relevant economic value in the crustacean business sector. Many factors as environmental pollution, catching/handling stressors or improper management of the cold chain may adversely affect quality and shelf-life of crustaceans. Fishing industry and trading companies are strongly requiring new scientific tools, as potential support in seafood management to get 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 an easy, potential source of nucleic acids. Despite their hypothesized degradation, we were able to detect specific nucleotide sequences. At NCBI database, we selected N. norvegicus mRNAs sequences as potential indicator(s) of molecular effects (stress/immune responses and/or degradations) of biological events linked to catching, handling, and freezing/thawing cycles. We obtained small amplicons of Actin, Cytochrome Oxidase I (COI), Calpain M, Prophenoloxidase (ppo) and Crustinlike antimicrobial peptide, in preliminary One Step RT-PCR using polyA+ RNA or total RNA. By comparative One Step RT-PCR and/or QRT-PCR assays we evaluated the amplification efficiency (AE) of the selected mRNAs using total RNA extracted from muscle exudates of thawed specimens, maintained at 4°C for 1, 4 and 7 days (shelf-life). We found that long shelf-life and lowest quality correlate with the lowest values of AE. Our results provide new opportunities in food science applications and in industrial-scale management, to track quality and shelf-life over time. 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.