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The microbial safety and stability of most food, are based on an application of preservative factors called hurdles. Each hurdle implies putting microorganisms in a hostile environment, which inhibits their growth or causes their death (Leistner, 2000). Some of those hurdles have been empirically used for years to stabilize meat, fish, milk and vegetables. This sometimes leads to completely different product with its own new taste characteristics. Examples of hurdles in marine products are salt (salted cod, klipfish), smoke (cold or hot smoked salmon, herring), acids (marinated products, pickles), temperature (high or low), fermentative microorganisms (traditional Asian sauces) and more recently redox potential (vacuum-packed products). Those preservative factors have been studied for years, but a large amount of potential hurdles for food have already been described including organic acids, bacteriocins, chitosan, nitrate, lactoperoxidase, essential oil, modified atmosphere packaging… , as well as novel decontamination technologies such as microwave and radio frequency, ohmic and inductive heating, high pressure, pulsed electric field, high voltage arc discharge, pulsed light, oscillation magnetic field, ultraviolet light, ultrasound, X-ray, electrolyse NaCl water, ozone… (Kim et al, 1999 ; Weber, 2000 ; Mahmoud et al, 2006). Hurdles that have a positive effect by inhibiting microorganisms may have a negative one on other parameters such as nutritional properties or sensory quality, depending on their intensity. As an example, salt content in food must be high enough to inhibit pathogens and spoilage microorganisms, but not so high to impair taste. In order to lower the preservative level, the hurdle technology concept has been developed (Leistner, 1985), consisting in using combined hurdles to establish an additive antimicrobial effect, and even sometimes a synergetic one, thus improving the safety and the sensory quality of food.
Various studies have shown that two-phase slurry ice is more efficient than ordinary flake ice for chilling fresh fish. In most of the studies only one type of slurry ice has been applied, most often prepared in commercial ice-machines. The objective of this work was to investigate both chilling and maintenance of low temperature utilising flake ice and different kinds of ice slurries, both from commercial ice-machines and also manually prepared by mixing crushed ice and brine. The slurry ice particles are smaller when produced in the ice machines than in the latter method and this small size of the ice particles is widely accepted as one of the predominant factors for rapid cooling of fish. Both saithe and a cylinder made of agar were used as specimen in the experiments. As in other similar studies the cooling rate of all of the different slurry ice types was superior compared to flake ice. Very similar cooling rates were gained using different ice slurries of the same temperature. Therefore, the most important property of the chilling medium was concluded to be temperature since the size of the ice particles seemed to have only minor influence on the cooling rate. The importance of distributing the ice medium evenly when packing fish and ice medium in fish tubs became evident in this work. In order to maintain low temperature during storage, ice slurries are only better than ordinary flake ice for the first few days of storage. After a few days the faster melting of the ice slurries results in inferior cooling capacity so the flake ice, in general, maintains lower temperature in fish through long storage. 1 Introduction Numerous papers showing comparison of chilling of fish with flake ice vs. chilling of fish with two-phase ice slurries have been published. Figure 1 shows the results of an experiment in which a 350 L insulated container was half filled with slurry ice and 30 kg of small to medium sized cod (Davies, 2005). The slurry ice was made of 3 wt. % salt brine with an ice fraction of 35 wt. % and a temperature of-2.6 °C. A second container was packed with cod and flake ice and the temperature evolution in the centre of fish specimen was recorded as in the first container. The performance of the slurry ice is obviously much better. However, the initial temperature of the flake iced fish actually seems to have been higher in the flake iced fish than in the slurry iced fish. More rapid cooling and higher heat transfer rates are explained by larger contact area between the fish and ice particles but also by the fact that the temperature difference between the chilling medium and the fish is larger in the slurry ice case. Similar studies have been done with plaice (Paul, 1998). The fish was cooled in boxes and the results, presented in Figure 2, show that the time required to chill the plaice below 2 °C was more than three times shorter for the slurry ice than for the flake ice. Other similar studies (e.g. Egolf el at., 2005) also show a superior cooling rate of slurry ice compared to flake ice for chilling fish. Figure 1. Cooling of cod using flake ice and slurry ice (Davies, 2005). Figure 2. Cooling of plaice using flake ice and slurry ice (Paul, 1998). Comparison between different slurry ice-and liquid ice types was not found in the literature. Neither has much been published about the cooling capacity of the different ice media. Therefore it was considered useful to include more than one type of slurry ice in the model studies in the present study and not only investigate cooling, but also the maintenance of low temperature during storage.
Bismuth compounds are widely used in industrial processes and products. In medicine, bismuth salts have been applied in combination with antibiotics for the treatment of Helicobacter pylori infections, for the prevention of diarrhea, and in radioimmunotherapy. In the environment, bismuth ions can be biotransformed to the volatile bismuth compound trimethylbismuth (Me3Bi) by methanobacteria. Preliminary in-house studies have indicated that bismuth ions are methylated in the human colon by intestinal microflora following ingestion of bismuth-containing salts. Information concerning cyto- and genotoxicity of these biomethylated products is limited. In the present study, we investigated the cellular uptake of an organic bismuth compound [monomethylbismuth(III), MeBi(III)] and two other bismuth compounds [bismuth citrate (Bi-Cit) and bismuth glutathione (Bi-GS)] in human hepatocytes, lymphocytes, and erythrocytes using ICP-MS. We also analyzed the cyto- and genotoxic effects of these compounds to investigate their toxic potential. Our results show that the methylbismuth compound was better taken up by the cells than Bi-Cit and Bi-GS. All intracellularly detected bismuth compounds were located in the cytosol of the cells. MeBi(III) was best taken up by erythrocytes (36%), followed by lymphocytes (17%) and hepatocytes (0.04%). Erythrocytes and hepatocytes were more susceptible to MeBi(III) exposure than lymphocytes. Cytotoxic effects of MeBi(III) were detectable in erythrocytes at concentrations >4 µM, in hepatocytes at >130 µM, and in lymphocytes at >430 µM after 24 h of exposure. Cytotoxic effects for Bi-Cit and Bi-GS were much lower or not detectable in the used cell lines up to a tested concentration of 500 µM. Exposure of lymphocytes to MeBi(III) (250 µM for 1 h and 25 µM/50 µM for 24 h) resulted in significantly increased frequencies of chromosomal aberrations (CA) and sister chromatid exchanges (SCE), whereas Bi-Cit and Bi-GS induced neither CA nor SCE. Our study also showed an intracellular production of free radicals caused by MeBi(III) in hepatocytes but not in lymphocytes. These data suggest that biomethylation of bismuth ions by the intestinal microflora of the human colon leads to an increase in the toxicity of the primary bismuth salt.
Antioxidative activity of hydrolyzed protein prepared from alkali-solubilized catfish protein isolates was studied. The isolates were hydrolyzed to 5, 15, and 30% degree of hydrolysis using the protease enzyme, Protamex. Hydrolyzed protein was separated into hydrolysates and soluble supernatants, and both of these fractions were studied for their metal chelating ability, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability, ferric reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC), and their ability to inhibit the formation of thiobarbituric acid reactive substances (TBARS) in washed tilapia muscle containing tilapia hemolysate. Both hydrolysates and supernatants were characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Results showed that DPPH radical scavenging ability and reducing power of catfish protein hydrolysates decreased, whereas the ORAC value, metal chelating ability, and ability to inhibit TBARS increased, with an increase in the degree of hydrolysis. Hydrolysate samples showed higher DPPH radical scavenging ability and Fe3+ reducing ability, and supernatant samples had higher metal chelating ability. In general, low molecular weight (MW) peptides had high ORAC values and high metal chelating ability, and high MW peptides had a higher reducing power (FRAP) and were more effective in scavenging DPPH radicals. In a washed muscle model system, the ability of catfish protein hydrolysates and their corresponding supernatants to inhibit the formation of TBARS increased with an increase in the degree of hydrolysis.
Monoheme cytochromes of the C-type are involved in a large number of electron transfer processes, which play an essential role in multiple pathways, such as respiratory chains, either aerobic or anaerobic, and the photosynthetic electron transport chains. This study reports the biochemical characterization and the crystallographic structure, at 1.23 Å resolution, of a monoheme cytochrome c from the thermohalophilic bacterium Rhodothermus marinus. In addition to an α-helical core folded around the heme, common for this type of cytochrome, the X-ray structure reveals one unusual α-helix and a unique N-terminal extension, which wraps around the back of the molecule. Based on a thorough structural and amino acid sequence comparison, we propose R. marinus cytochrome c as the first characterized member of a new class of C-type cytochromes.
Enzymatically hydrolyzed fish protein hydrolysates could be used as a source of antioxidative nutraceuticals. In our current work, we have investigated alkali-solubilized tilapia (Oreochromis niloticus) protein hydrolysates for their ability to scavenge reactive oxygen species (ROS) and for their reducing power. Tilapia protein isolate was prepared by an alkaline solubilization technique and used as a substrate for enzyme hydrolysis. Cryotin, protease A ‘Amano′ 2, protease N ‘Amano′, Neutrase and Flavourzyme, were used separately to determine their effectiveness in hydrolyzing tilapia protein isolate. ROS scavenging ability was quantified using an isoluminol enhanced chemiluminescent assay in the presence of a) hydrogen peroxide or b) mononuclear cells isolated from human blood. Ferric reducing antioxidant power (FRAP) and Trolox equivalent antioxidant capacity (TEAC) of the hydrolysates using 2, 2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) or 2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), were also investigated. Results showed that, in general, the TEAC, FRAP values and ROS scavenging ability of the hydrolysates increased with an increase in the degree of hydrolysis. Among the different hydrolysates, those prepared using Cryotin were most effective and Amano A2 hydrolysates were least effective in scavenging ABTS·+ and ROS generated by hydrogen peroxide. However, FRAP assay showed that hydrolysates prepared using Flavourzyme were most effective, and Amano N and Neutrase hydrolysates were least effective in reducing ferric ions. No significant difference was observed among the hydrolysates produced with different enzymes in their ability to scavenge ROS generated by phorbol myristate acetate stimulated mononuclear cells. These results shed light on the in vitro ROS scavenging ability of alkali solubilized tilapia protein hydrolysates, as well as potential nutraceutical use of these hydrolysates.
Eighteen new microsatellite loci consisting of 10 di-, 5 tri-, 2 tetra- and 1 heptanucleotide repeats are introduced for the Atlantic cod (Gadus morhua L.). All loci were co-amplified in two polymerase chain reactions (plus two previously published microsatellites) and all products were typed clearly. The number of alleles per locus ranged from six (PGmo130) to 45 (PGmo76) and the observed heterozygosity ranged from 0.356 (PGmo130) to 0.957 (PGmo95). All loci except one followed Hardy–Weinberg expectations. Genetic linkage disequilibrium analysis between all pairs of loci did not yield any significant values.
Changes in the conformation of catfish (Ictalurus punctatus) myosin due to (i) cations (ii) alkaline pH and (iii) salt addition were determined using circular dichroism, tryptophan fluorescence, differential scanning calorimetry and hydrophobicity studies. The relation between conformation and storage modulus (G′) of alkali treated myosin was studied. Two types of bases, NaOH and KOH were used for unfolding myosin under three alkaline conditions, pH 11.0, 11.5 and 12.0. Myosin, unfolded under alkali conditions was immediately refolded by adjusting pH back to 7.3. Subjecting myosin to alkaline conditions and subsequent readjustment to pH 7.3 increased the G′ of thermally treated myosin. G′ was affected by the presence or absence of salt during alkali treatments. When salt was present during alkali unfolding of myosin, the added salt stabilized the conformation of myosin against alkali unfolding and denaturation. In the absence of salt or when salt was added after refolding, myosin showed significantly higher denaturation and high G′ on heating and cooling. Among the different alkaline pH values, myosin treated at pH 11.0 showed higher G′. The type of anions influenced the conformation of myosin and the strength of gels. Treatment of myosin with KOH resulted in greater denaturation and higher gelling ability (G′) compared to NaOH.