Blue-back fish: Fatty acid profile in selected seasons and retention upon baking

The effects of the catching season (either Autumn/Winter or Spring) on lipid content, fatty acid profile and true retention values after oven baking were determined in anchovy (Engraulis encrasicholus), sardine (Sardina pilchardus), sprat (Sprattus sprattus) and horse mackerel (Trachurus trachurus) from the North Adriatic sea. In the raw state, the catching season induced significant changes in the flesh lipid contents of sardine and sprat. Anchovy was the species whose fatty acid composition of flesh lipids was most clearly affected by the season of catch. Oven baking had a significant, though rather modest, effect on some fatty acids and indices of sardine and anchovy lipids. When compared to the other species within the same season, spring sardine and winter sprat gave significantly higher true retention values for several individual fatty acids and some sums. Between 1 and 2.5 servings/week (depending on season) of either cooked anchovy, sardine, sprat, or 3 servings of cooked spring horse mackerel would suffice to satisfy human weekly requirements of EPA + DHA.     

Purification and characterization of trypsin from the viscera of sardine (Sardina pilchardus)

Trypsin from the viscera of Sardina pilchardus was purified by fractionation with ammonium sulphate, heat treatment and Sephadex G-100 gel filtration with a ninefold increase in specific activity and 9% recovery. The molecular weight of the enzyme was estimated to be 25,000 Da on SDS–PAGE. This enzyme showed esterase-specific activity on Nα-benzoyl-l-arginine ethyl ester (BAEE). The purified enzyme was inhibited by benzamidine, a synthetic trypsin inhibitor, and phenylmethylsulphonyl fluoride (PMSF) a serine-protease inhibitor, but was not inhibited by the β-mercaptoethanol. The optimum pH and temperature for the enzyme activity were pH 8.0 and 60 °C, respectively. The relative activity at pH 9.0 was 95.5% and the enzyme showed pH stability between 6.0 and 9.0. The N-terminal amino acid sequence of the first 12 amino acids of the purified trypsin was IVGGYECQKYSQ. S. pilchardus trypsin, which showed high homology to other fish trypsins, had a charged Lys residue at position 9, where Pro or Ala are common in fish trypsins. The enzyme was strongly inhibited by Zn²⁺ and Cu²⁺.     

Partial characterisation of gelatinolytic activities in herring (Clupea harengus) and sardine (Sardina pilchardus) possibly involved in post-mortem autolysis of ventral muscle

The present work aims at identifying enzymatic activities that may contribute to the post-mortem autolysis of the ventral muscle, known as belly bursting, in herring (Clupea harengus). Gelatinolytic proteases extracted from several herring tissues and also from a sardine (Sardina pilchardus) tissue were partially characterised using gelatin zymography, inhibitor analysis, immunodetection with anti-trypsin antibody and MALDI-TOF/TOF peptide sequencing. The results indicate the presence of gelatinolytic trypsin-like serine proteases and metalloproteases in several samples including the ventral muscle of herring. MS/MS analysis gave partial sequences of peptides from some of the proteases, and these were identified as elastase, trypsin and aspartyl aminopeptidase. It is likely that belly bursting in herring is caused by leakage of enzyme-rich fluids from the intestine and/or pyloric caeca which may also contain exogenous proteases from the digestive systems of the prey.     

Physical,chemical and sensory analysis of sardine (Sardina pilchardus) stored in ice

The shelf-life of iced sardine (Sardina pilchardus) was studied. The main changes which take place in raw fish were investigated by means of organoleptic assessments, chemical analyses (total volatile nitrogen (TVB-N), trimethylamine (TMA-N), trimethylamine oxide (OTMA-N) and hypoxanthine) and physical measurements (GR Torrymeter readings and pH). The influence of both seasonal changes and fat deterioration were also considered. The results obtained indicate that TVB-N and TMA-N parameters are not good freshness indicators for this species, but Torrymeter readings and hypoxanthine values can be used as indicators of freshness. However, they must always be confirmed by a sensory evaluation. From the different combinations tried, the most highly significant degree of correlation was obtained between sensory evaluation and Torrymeter readings.     

Differential detection of small pelagic fish in Tunisian canned products by PCR-RFLP: An efficient tool to control the label information

Correct identification of fish species including their origin requires a compilation of recognized material to verify the conformity of the product with the labelling requirement. Sardine is among the internationally disputed example of species identification conflicts, with the lack of genetic reference material at the Southern part of the Mediterranean. Therefore a method was developed to discriminate between Tunisian small pelagic fish: Sardina pilchardus, Sardinella aurita and Engraulis encrasicolus. DNA extraction from fresh fish and from 12 canned sardine and 2 anchovy-type products, was followed by a PCR method that specifically amplifies a 252 bp fragment of the cytochrome b gene. The new sequences which were deposited in the NCBI GenBank, were searched against a nucleotide sequence database (GenBank) and phylogenetically analyzed with the MEGA software. Multiple alignments of 3 analyzed reference samples belonging to Clupeomorpha species was performed versus the canned samples. Low intraspecific variability was observed for canned anchovy and sardine (<0.05), whereas mean interspecific variability was 0.23. A phylogenetic tree was constructed, and the calculated bootstrap values (BP, 71-100%) were used as indicators of the correct assignment of unknown canned samples to reference species. Postamplification digestion with HaeIII and ALuI restriction enzymes, yielded specific profiles that enabled anchovy to be distinguished from either of the sardine species. This PCR-RFLP was found to be reliable for species identification of canned fish products.     

Use of eye fluid refractive index in sardine (<Emphasis Type="Italic">Sardina pilchardus</Emphasis>) as a freshness indicator

In this study the usage of eye fluid refractive index as a freshness indicator for sardine (Sardina pilchardus) was investigated. Eye fluid refractive index measurements and quality control analyses of sardine during storage at 0°C and +4°C were performed at 24h intervals. According to the sensory analysis results, the sardines stored at 0°C and +4°C had a shelf life of 6 and 4days, respectively. The changes in eye fluid refractive index values during storage at 0°C were not significant while the changes at +4°C were significant (p<0.01). TVB-N and TMA-N values significantly (p<0.01) increased. No microbiological growth was observed at 0°C, however the increase in microorganism counts was significant at +4°C. As a result, eye fluid refractive index measurements can be used as a quality criterion for sardine freshness, when stored at +4°C.     

Drying of sardine muscles: Experimental and mathematical investigations

The aim of this work was to study the effect of air drying process on the dehydration kinetics of sardine muscles (Sardina pilchardus). Experimental drying kinetics were measured at five air temperatures (40, 50, 60, 70 and 80 °C), two relative humidity and at a constant air velocity of 1.5 m/s. The sardine drying kinetics were accelerated by increasing air temperature and were showed down when increasing air humidity. Moisture desorption isotherms of sardine muscles were determined at three temperatures (40, 50 and 70 °C) by using the static gravimetric method. The equilibrium moisture contents of sardine muscles were used to treat mathematically the experimental drying kinetics. Experimental drying kinetics and desorption isotherms of sardine muscles were described by using empiric models available in the literature. Eight models (GAB, BET, Henderson–Thompson, Modified Chung & Pfost, Modified Halsey, Oswin, Peleg and Adam & Shove models) were compared in order to describe the desorption isotherms. The Peleg model showed the best fitting of experimental data. For the drying kinetics, the Page model allowed a better fitting than the Newton and the Henderson and Pabis models. The Page model was thus used for simulating the drying kinetics of sardine muscles between 40 and 80 °C.