The antioxidant activities of alkali-treated tilapia protein hydrolysates were determined by their ability to inhibit the formation of lipid hydroperoxides (PV) and thiobarbituric acid reactive substances (TBARS) in a washed muscle model system and by their ability to inhibit DPPH free radicals and chelate ferrous ion in an aqueous solution. Protein isolates were prepared from tilapia white muscle using alkali solubilization at pH 11.0 and reprecipitation at pH 5.5. Protein hydrolysates were prepared by hydrolyzing the isolates using five different enzymes, Cryotin F, Protease A Amano, Protease N Amano, Flavourzyme, and Neutrase, to 7.5, 15, and 25% degrees of hydrolysis (DH). All of the protein hydrolysates significantly (p < 0.05) inhibited the development of TBARS and PV. The antioxidant activity of the hydrolysates increased with the DH. Also, the antioxidant activity of the hydrolysates varied significantly (p < 0.05) among the different enzymes. The ability of different enzyme-catalyzed protein hydrolysates to scavenge DPPH radicals was not reflected in their ability to inhibit oxidation in a washed tilapia model system. In a washed muscle model system, the hydrolysates prepared using Cryotin F were most effective and the hydrolysates prepared using Flavourzyme and Neutrase were least effective in inhibiting the development of TBARS and PV, whereas in an aqueous solution, hydrolysates prepared using Flavourzyme were most effective in scavenging DPPH radicals and chelating ferrous ions. Enzymatic hydrolysis decreased the size of tilapia protein hydrolysates and, in general, tilapia protein hydrolysates with low molecular weights were better antioxidants than those with high molecular weights.
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BACKGROUND: Color of muscle foods plays a major role in consumer perception of meat quality. Carbon monoxide (CO) has been successfully used for improving color of packaged meat and fish products. In this study, we wanted to investigate pre-mortem treatment of live tilapia using 100% CO for its ability to improve the color of frozen whole tilapia. We compared untreated and CO-treated whole, gutted tilapia, frozen for 2 and 4 months at − 20 °C. Frozen tilapia samples were thawed overnight at 4 °C, filleted and analyzed for their color, heme peak wavelength and CO concentration.
RESULTS: Euthanasia using CO significantly increased redness (a* value) and lightness (L* value) of tilapia white and red muscle. Frozen storage significantly (P < 0.05) decreased redness of both CO-treated and untreated tilapia. However, even after 4 months of frozen storage, a*-value of CO-treated tilapia was similar to fresh untreated tilapia fillets. Heme peak wavelengths of CO-euthanized tilapia were higher than in untreated tilapia and there was no significant (P > 0.05) decrease in heme peak wavelengths of CO-treated tilapia white and red muscle during frozen storage. The CO content of frozen euthanized tilapia fillets was significantly (P > 0.05) higher than in untreated fillets. In general, red muscle tissue of euthanized tilapia had a higher concentration of CO than white muscle.
CONCLUSION: Color stability of tilapia fillets was significantly improved by pre-mortem CO treatment. The color of CO-treated fillets was retained during frozen storage compared to untreated fillets. Hence, pre-mortem CO treatment could be used as a new method for improving color of tilapia. Copyright © 2008 Society of Chemical Industry
Carbon monoxide (CO) has been used for improving the color of muscle foods. In the current study, we compared the postmortem treatment of tilapia fillets with 100% CO and euthanasia of live tilapia with CO for their ability to stabilize the color of white and red muscle of tilapia fillets. Both postmortem CO treatment and CO euthanasia were effective in increasing the redness (a* value) and lightness (L* value) of tilapia white and red muscle. Fillets obtained from CO-euthanized tilapia showed significantly higher a* and L* values during 1 mo of frozen storage at –20 °C and subsequent thawing and storage at 4 °C for 18 d. The amount of CO present in the red and white muscles decreased during the 18 d of storage at 4 °C. There was no significant difference in the pH, drip, or thaw loss of CO-treated tilapia fillets compared to the untreated fillets.
Green, nonsulfur-like bacteria (GNSLB) and cyanobacteria form major components of microbial mats in both sulfidic and non-sulfidic hot springs and have been mainly studied in hot springs of Yellowstone National Park (YNP). These organisms synthesize specific lipid biomarkers such as wax esters and long chain polyunsaturated alkenes (GNSLB) and heptadecane (cyanobacteria). We analyzed the lipid distribution and their stable carbon isotopic composition in sulfidic Icelandic hot spring microbial mats known to contain GNSLB and cyanobacteria. Based on the lipid distribution, it seems that the GNSLB in these mats are closely related to Chloroflexus aurantiacus. The stable carbon isotopic composition of the bulk biomass and wax esters suggests mainly autotrophic growth by GNSLB in this sulfidic hot spring. However, the stable carbon isotopic composition of hentriacontatriene in the two GNSLB mats suggests an alternative carbon source for the C31:3 alkene producing GNSLB from that in YNP. The isotopic composition of cyanobacterial biomarkers in the mat most distant from the source of the hot spring seems to suggest inorganic carbon limitation for cyanobacteria, possibly because they grow underneath the GNSLB in these sulfidic hot spring inverted microbial mats.
Starch and pullulan-modifying enzymes of the α-amylase family (glycoside hydrolase family 13) have several industrial applications. To date, most of these enzymes have been derived from isolated organisms. To increase the number of members of this enzyme family, in particular of the thermophilic representatives, we have applied a consensus primer-based approach using DNA from enrichments from geothermal habitats. With this approach, we succeeded in isolating three new enzymes: a neopullulanase and two cyclodextrinases. Both cyclodextrinases displayed significant maltogenic amylase side activity, while one showed significant neopullulanase side activity. Specific motifs and domains that correlated with enzymatic activities were identified; e.g., the presence of the N domain was correlated with cyclodextrinase activity. The enzymes exhibited stability under thermophilic conditions and showed features appropriate for biotechnological applications.
The effect of filtered wood smoke (FS) processing on microbial growth in yellowfin tuna steaks was investigated. Steaks were treated for 32 h and then subjected to aerobic storage for 8 days and levels of aerobic bacteria studied. The FS treatment was compared to untreated tuna (exposed to air), 100% nitrogen (N2) (to investigate the specific effect of oxygen exclusion), 100% carbon monoxide (CO) (to investigate the effect of the CO molecule) and artificial FS (to investigate a gas with all the same gas compounds except for the smoke compounds). A study was also conducted with 21% carbon dioxide (CO2) to investigate the effect of CO2, which is found in both FS and the artificial FS. The results show that all treatments effectively (p<0.05) reduced aerobic microorganisms compared to the untreated control after 32 h, possibly due to an oxygen exclusion effect as well as to the presence of other active compounds in the gases. However, only FS, artificial FS, 100% CO and 21% CO2 treatments had a significant (p>0.05) residual antibacterial/bacteriostatic effect post-treatment. The FS treatment had the most significant residual effect (p<0.05), and this study strongly suggests this effect is due to the presence of CO2, smoke components and CO.
Survival problems are encountered at early stages of intensive fish rearing and antibiotics are widely used to remedy the situation. Probiotics may provide a potential alternative method to protect larvae from opportunistic and pathogenic bacteria and promote a balanced environment. This study was designed to search for new probiotics to target this critical period in cod rearing. Potential probionts were selected from the natural microbiota of cod aquacultural environment. The selection was based on several criteria: pathogen inhibition potential, growth characteristics, strain identification, metabolite production and adhesion to fish cell lines. Our study demonstrated that 14% of screened bacteria (n = 188) had antagonistic properties towards fish pathogens. The majority of these isolates were Gram-positive (81%), belonging to Firmicutes (69.2%) and Actinobacteria (11.5%) phyla based on 16S rRNA gene sequencing. Only 6 (3.2%) of 188 isolates could inhibit all three pathogens tested: Vibrio anguillarum, Aeromonas salmonicida subsp. achromogenes and Vibrio salmonicida. Differences observed in activity intensity and spectrum among inhibitory isolates emphasise the need to develop probiotic mixtures for efficient prophylactic methods. Comparison of growth behaviour of inhibitory isolates and pathogens at cod rearing temperatures, metabolite production and adhesion capacity were considered for final probiont selection. Four promising isolates that could be used as a mixed supplement to rearing water were identified as putative probiotic bacteria. This study emphasises the importance and potential of lactic acid bacteria in aquaculture.
A speciation technique for arsenic (As) has been applied using ion pair reverse phase-high performance liquid chromatography coupled to inductively coupled plasma mass spectrometry (RP-HPLC-ICP-MS). Six As-species (arsenite, arsenate, dimethylarsinic acid, dimethylarsinous acid, monomethylarsonic acid and monomethylarsonous acid) have been separated with isocratic elution within less than 6 minutes. A cation exchange column was used for separation of AsB, AsC, Tetra (Me4As+) and TMAsO. The As chemical form and oxidation state is highly important in respect to toxicity; therefore total As-determination is insufficient for a complete toxicological evaluation and risk assessment. Arsenic-speciation has been studied on urine samples of children from an As-affected area in the Iron Quadrangle, Brazil. DMAsV and MMAsV were the major urinary metabolites in these samples. The mean value for total As-concentration of all samples (n = 15) was 26.3 ng As mL−1 with a range of 16.1 to 55.2 ng As mL−1. TMAsO and AsC (arsenocholine) were not detected in this study. In most samples, monomethylarsonous acid [MMAsIII] was detected up to 2.0 ng As mL−1. DMAsIII was not detected at any time, most probably due to volatilisation and some oxidation to DMAsV. The chromatographic recoveries, calculated from ([sum(species) × 100]/total As in urine samples), ranged from 77.4 to 94.9%. This work also contributes to the pertinent discussion regarding the reliability of MMAsIII/DMAsIII speciation in urine.
A bit fishy: Six arsenolipids have been isolated from cod-liver oil and identified by HPLC and mass spectrometry as a series of arsenic-containing long-chain fatty acids (for example, see picture). These fatty acids account for about 20 % of the total arsenolipid content of cod-liver oil.
Epidemiological studies have indicated that exposure of humans to inorganic arsenic in drinking water is associated with the occurrence of bladder cancer. The mechanisms by which arsenic induces this malignancy are still uncertain; however, arsenic metabolites are suspected to play a pivotal role. The aim of the present study was the investigation of uptake capabilities of human urothelial cells (UROtsa) compared with primary human hepatocytes (phH) as well as the intracellular distribution of the arsenic species. Additionally, we were interested in the cyto- and genotoxic potential (comet assay, radical generation) of the different arsenic compounds in these two cell types. Our results show that UROtsa cells accumulate higher amounts of the arsenic species than the phH. Differential centrifugation revealed that the arsenic compounds are preferentially distributed into nuclei and ribosomes. After 24-h exposure, arsenic is mainly found in the ribosomes of UROtsa cells and in the nuclei and mitochondria of phH. In contrast to the pentavalent arsenic species, the trivalent species induced a 4- to 5-fold increase of DNA damage in hepatocytes. Radical generation, measured by thiobarbituric acid reactive substances, was more pronounced in hepatocytes than in urothelial cells. In summary, the uptake of arsenic compounds appears to be highly dependent upon cell type and arsenic species. The nonmethylating urothelial cells accumulate higher amounts of arsenic species than the methylating hepatocytes. However, cyto- and genotoxic effects are more distinct in hepatocytes. Further studies are needed to define the implications of the observed accumulation in cellular organelles for the carcinogenic activity of arsenic.