Kateřina Malachová

The use of the fungus Dichomitus squalens for degradation in rotating biological contactor conditions.

Biodegradation potential of Dichomitus squalens in biofilm cultures and rotating biological contactor (RBC) was investigated. The fungus formed thick biofilms on inert and lignocellulosic supports and exhibited stable activities of laccase and manganese peroxidase to reach 40-62 and 25-32% decolorization of anthraquinone Remazol Brilliant Blue R and heterocyclic phthalocyanine dyes, respectively. The decolorization ceased when glucose concentration dropped to 1 mmol l(-1). In RBC reactor, respective decolorizations of Remazol Brilliant Blue R and heterocyclic Methylene Blue and Azure B dyes (50 mg l(-1)) attained 99%, 93%, and 59% within 7, 40 and 200 h. The fungus exhibited tolerance to coliform and non-coliform bacteria on rich organic media, the inhibition occurred only on media containing tryptone and NaCl. The degradation efficiency in RBC reactor, capability to decolorize a wide range of dye structures and tolerance to bacterial stress make D. squalens an organism applicable to remediation of textile wastewaters.

Anti)mutagenic and immunomodulatory properties of quercetin glycosides

BACKGROUND Quercetin-3-O-β-D-glucopyranoside (isoquercitrin) and quercetin-3-O-rutinoside (rutin) are common components of a normal human diet and are increasingly used in food supplements. Here their effect on mutagenesis and immunity is shown. RESULTS The in vitro (anti)mutagenic potential was compared with that of quercetin using the Ames test in Salmonella typhimurium His(-) strains TA100, TA98 and TA102. Isoquercitrin only slightly increased the number of revertants, while rutin was totally non-mutagenic. On the other hand, all compounds displayed dose-dependent protective activity against H2O2 - and tert-butyl hydroperoxide-induced oxidative damage to the TA102 strain and at 75 µmol L(-1) inhibited H2O2/Fe(2+)-induced formation of the open circular and linear forms of the DNA plasmid pBSIISK(-). In mice, none of the flavonols (0.86 µmol day(-1), 34 days) induced harmful effects. In immunized animals, all compounds enhanced ex vivo B cell proliferation; quercetin stimulated lymphocyte basal proliferation and increased the number of IgM-producing lymphocytes. Rutin promoted NK cytotoxic activity, supported T cells and enhanced gut epithelium renewal. No effect on IgG-forming cells was found. CONCLUSION Isoquercitrin displayed negligible and rutin no mutagenicity, but both showed significant antimutagenic and DNA-protective effects against oxidative damage. In vivo, they supported the readiness of the immune system for specific humoral immune response.

Effect of bacteria on the degradation ability of Pleurotus ostreatus

White-rot fungi are efficient degraders of lignin whose extracellular enzymes have a potential to degrade organopollutants. In natural conditions these fungi enter into interactions with other organisms, which may affect their biodegradation capacity. The aim was to investigate the ability of Pleurotus ostreatus to form stable biofilms and to test the capacity of the fungus to degrade Remazol Brilliant Blue R in mixed cultures with bacteria. Bacterial counts were determined to see the behavior of the bacterium in the mixed culture with the fungus. In axenic conditions, the homogenized fungal mycelium was able to form an active biofilm which quickly degraded the dye. The addition of Pseudomonas fluorescens or Bacillus licheniformis bacteria at 106CFU·mL-1 did not affect the decolorization rate by 7-d-old fungal biofilms where the decolorization rate reached 90%. In contrast, when fragments of the fungal mycelium were used for inoculation to pre-formed biofilm of P. fluorescens, the biofilm was allowed to develop for one weeks time, no decolorization of RBBR was observed and low activities of MnP and laccase were detected. The use of agar disks covered with fungal mycelium for the inoculation to pre-formed biofilm of P. fluorescens resulted in a fully developed biofilm that decolorized RBBR with similar efficiency as the pure P. ostreatus. The difference between the agar-disk- and homogenized-mycelium inoculated fungal biofilms was corroborated by the measurement of total fungal biofilm biomass that was 6-fold lower in the latter biofilm. Capability of the fungus to overcome the competition of the bacterial biofilm thus depended on the type of fungal growth centres, where intact hyphae were superior to the fragments of mycelium. A similar effect was not observed with the biofilms of B. licheniformis where the bacterial growth was less massive. The ability of P. ostreatus biofilms to resist massive bacterial stress was demonstrated.

Effect of yeasts on biodegradation potential of immobilized cultures of white rot fungi

The aim was to investigate the effect of yeast organisms on the degradation process by immobilized cultures of ligninolytic fungi. Immobilization was accomplished by 7-day colonization of polyamide mesh with mycelial fragments. Irpex lacteus decolorized >90% of the initial concentration of 150mgl-1 of anthraquinone Remazol Brilliant Blue R dye in three subsequent decolorization cycles and the degradation capacity was not negatively affected by the presence of 106Saccharomyces cerevisiae cells per ml in the mixed culture. The yeast was not able to degrade the dye. I. lacteus biofilm was also resistant to bacterial infection with E. coli. Inoculation of the yeast to pre-formed I. lacteus biofilm culture resulted in a reduction of fungal biomass by 27%. Levels of LiP, MnP and laccase of I. lacteus were not much influenced by S. cerevisiae or E. coli. Similar resilience of P. ostreatus biofilms was observed after exposure to yeast Issatchenkia occidentalis when the fungal degradation capacity measured with Reactive Orange 16 azo dye was maintained over two decolorization cycles. I. occidentalis did not degrade the dye under the conditions used. Formation of densely packed fungal biofilms with abundant extracellular polysaccharide was not impeded by the yeast. Increase of MnP and laccase levels attributable to the presence of I. occidentalis was observed.

Genotoxicity Estimation in Soils Contaminated with Polycyclic Aromatic Hydrocarbons after Biodegradation

Polycyclic aromatic hydrocarbons (PAHs) belong to recalcitrant pollutants that resist to decomposition by natural processes. As a result of long term industrial activities and accidents, PAHs and their residues accumulate in soil. Biodegradation processes that are able to decompose PAHs and their mixtures are rather complex and can be affected by many physical, chemical and biological factors [9, 5], Some PAHs are degraded to produce intermediates with a mutagenic activity, such as dihydrodiols, phenols, arene oxides, etc. [3, 1]. The purpose of the study was to detect mutagene in soil contaminated with coke plant wastes in the course of many years of production and, on the basis of genotoxicity changes, evaluate the efficiency of biodegradation technologies used for decontamination.

Antigenotoxic effects of a bark extract from Magnolia officinalis

Abstract The aim was to detect antimutagenic and DNA protective effects of a plant extract from bark of Magnolia officinalis adversing oxidative DNA damage. The ability to inhibit mutagenicity induced by tert-butyl hydroperoxide (t-BOOH) and hydrogen peroxide (H2O2) was determined with Ames test using Salmonella typhimurium His- TA102 bacterial strain. Inhibition values of 72.8 and 98.7 % were detected for t-BOOH and H2O2, respectively. The protective effect of the extract against DNA strand scission induced by hydroxyl radicals was studied with plasmid pBluescript II SK(-). The analysis of DNA strand breaks in the plasmid DNA proved a significant inhibition of DNA damage.