The control of the microbiological quality of bivalve molluscs assumes particular importance because they are among the most produced seafood products and mostly consumed as a whole, raw, or lightly cooked. The composition of the bacterial community associated with bivalves depends mostly on the microbiology of the surrounding environment at growing sites. Once the relationship between microbiology of bivalves and environment is established, a better classification and monitoring of the shellfish beds and evaluation of depuration strategies can be achieved. In this work, we tested if the methods of DNA extraction commonly used for the culture-independent microbiological analysis of sediment and water could be used directly, or with modifications, for the analysis of bacteria in mussels. The commercial kits Genomic DNA Purification Kit (MBI Fermentas, Vilnius, Lithuania), UltraCleanTM Soil DNA Isolation Kit (MOBIO Laboratories, Inc., Carlsbad, CA) and the method described by Griffiths and collaborators for DNA/RNA co-extraction were compared. The efficiency of extraction was assessed by DNA fluorescence and the denaturing gradient gel electrophoresis gel patterns of 16S ribosomal RNA gene fragments were used to compare the reproducibility and representativeness of the extraction methods. Results showed that the DNA/RNA co-extraction method with modifications was the most suitable. However, the results must be interpreted in the light of the purpose of the study and the relevance of maximizing extraction yield or diversity estimate, without compromising reproducibility. To our knowledge, this was the first attempt to transpose the procedure currently used for DNA extraction from sediments or waters, to the analysis of whole mussels. 相似文献
High molecular weight Maillard reaction products (melanoidins) are described to possess metal-chelating properties. Whereas
in food systems, this ability contributes to antioxidant properties, the consequences on biological systems are not quite
clear. The study was aimed to evaluate the implication of metal complexation by melanoidins on DNA damage. Melanoidins prepared
with d-glucose and different l-amino acids under water-free reaction conditions were charged with cupric ions. The effect on isolated DNA was investigated
by the PM2 assay and on cellular systems in the human colon carcinoma cell line HCT-116 by alkaline unwinding. Independent
of the amino acid composition, pure melanoidins (MW >14 kDa) did not cause significant DNA damage. By charging melanoidins
with Cu2+ ions, a considerable DNA strand breaking activity was detectable, which was again amplified in an oxidative milieu (addition
of hydrogen peroxide). Since Cu2+ normally does not provoke the formation of reactive oxygen species (ROS) via Fenton-type reaction, the results obtained have
to be attributed to reducing properties of melanoidins. Thus, in melanoidin–copper complexes redox cycling may take place
leading to Cu+ which subsequently undergoes Fenton-type and Haber–Weiss reactions. As a consequence, ROS are formed, which may explain the
generation of DNA strand breaks. 相似文献
The effect of γ-radiation on green onion DNA integrity, phenol content, oxygen radical absorbance capacity, employing pyrogallol red and fluorescein as probes, as well as ascorbic acid content has been evaluated. Measurements using thiazole orange-DNA fluorescence and agarose gel electrophoresis show that γ-radiation does not lead to an apparent DNA change in green onion. However, it was readily cleaved upon irradiation from the previously isolated nucleic acid. Furthermore, green onion exposure to γ-radiation produces slight increases in the polyphenol concentrations (163–188 μM Trolox eq.) and a decrease in the oxygen radical absorbance using fluorescein (from 245 to 200 Trolox eq.) Interestingly, a high ascorbic acid content (364 μM), which decreases by 40% after γ-ray exposure was measured by using pyrogallol-red-based oxygen radical absorbance capacity induction times from green onion aqueous extracts. Thus, our results suggest that ascorbic acid present in green onion plays a fundamental role in the plant antioxidant response toward γ-radiation exposure, while polyphenols remain largely unchanged, as revealed from oxygen radical absorbance capacity, employing pyrogallol red. 相似文献
This study was conducted to design a biosensor as a new, rapid, and sensitive tool for investigation of binding of zearalenone with double-stranded DNA (dsDNA). Polydiallyldimethylammonium chloride (PDDA) as a polycation and multiwall carbon nanotubes (MWCNTs) provide a positively charged surface with a high surface area for the immobilization of dsDNA as a polyanion on the surface of pencil graphite electrode (PGE). Using the dsDNA/MWCNT–PDDA-modified PGE, it was possible to detect the interaction of zearalenone with dsDNA, which allowed us to apply the dsDNA-modified electrode for trace determination of zearalenone. The changes at the oxidation signal of adenine were evaluated before/after each modification/immobilization step. By using dsDNA/PDDA–MWCNT/PGE, zearalenone could be detected as low as 0.005 ng mL?1. The relative standard deviation of five measurements of 0.5 ng mL?1 zearalenone was found to be 4.2 %. Finally, the highly stable electrochemical biosensor was applied to analyze the zearalenone concentration in milk and wheat samples.
