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33Hákarl is produced by curing of the Greenland shark (Somniosus microcephalus) flesh, which before fermentation is 34 toxic due to the high content of trimethylamine (TMA) or trimethylamine N-oxide (TMAO). Despite its long history of 35 consumption, little knowledge is available on the microbial consortia involved in the fermentation of this fish. In the 36 present study, a polyphasic approach based on both culturing and DNA-based techniques was adopted to gain insight 37 into the microbial species present in ready-to-eat hákarl. To this aim, samples of ready-to-eat hákarl were subjected to 38 viable counting on different selective growth media. The DNA directly extracted from the samples was further 39 subjected to Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) and 16S amplicon-40 based sequencing. Moreover, the presence of Shiga toxin-producing Escherichia coli (STEC) and Pseudomonas 41 aeruginosa was assessed via qualitative real-time PCR assays. pH values measured in the analyzed samples ranged 42 from between 8.07±0.06 and 8.76±0.00. Viable counts revealed the presence of total mesophilic aerobes, lactic acid 43 bacteria and Pseudomonadaceae. Regarding bacteria, PCR-DGGE analysis highlighted the dominance of close relatives 44 of Tissierella creatinophila. For amplicon sequencing, the main operational taxonomic units (OTUs) shared among the 45 data set were Tissierella, Pseudomonas, Oceanobacillus, Abyssivirga and Lactococcus. The presence of Pseudomonas 46 in the analyzed samples supports the hypothesis of a possible role of this microorganism on the detoxification of shark 47 meat from TMAO or TMA during fermentation. Several minor OTUs (<1%) were also detected, including 48 Alkalibacterium, Staphylococcus, Proteiniclasticum, Acinetobacter, Erysipelothrix, Anaerobacillus, Ochrobactrum, 49 Listeria and Photobacterium. Analysis of the yeast and filamentous fungi community composition by PCR-DGGE 50 revealed the presence of close relatives of Candida tropicalis, C. 52 tenuissimum, Moristroma quercinum and Phoma/Epicoccum, and some of these species probably play key roles in the 53 development of the sensory qualities of the end product. Finally, qualitative real-time PCR assays revealed the absence 54 of STEC and Pseudomonas aeruginosa in all of the analyzed samples. 55 56

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The possibility to clone, express and purify recombinant enzymes have originated the opportunity to dispose of a virtually infinite array of proteins that could be used in the clinics to treat several inherited and acquired pathological conditions. However, the direct administration of these recombinant proteins faces some intrinsic difficulties, such as degradation by circulating proteases and/or inactivation by the patient immune system. The use of drug delivery systems may overcome these limitations. Concerning recombinant enzyme therapy, the present review will mainly focus on the exploitation of erythrocytes as a carrier system for enzymes removing potentially noxious metabolites from the circulation, either as limiting treatment strategy for auxotrophic tumours or as a detoxing approach for some intoxication type inherited metabolic disorders. Moreover, the possibility of using RBCs as a potential delivering system addressing the enzymes to the monocyte-macrophages of reticular endothelial system for the treatment of diseases associated with this cell lineage, e.g. lysosome storage diseases, will be briefly discussed.