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Study of the interaction between fungus Pleurotus ostreatus and bacterial cultures on the abiotic surfaces - morphological, biochemical and proteomic analysis
Kozická, Barbora ; Petráčková, Denisa (advisor) ; Konopásek, Ivo (referee)
Ligninolytic fungi are well known for their ability to degrade a wide range of xenobiotics contaminating the environment, including synthetic industrial dyes. In this work Pleurotus ostreatus was used for decolorization of a synthetic textile dye Remazol Brilliant Blue R (RBBR). To set up a model fungal "fixed-bed" bioreactor the fungus was immobilized on a polyurethane foam and artificially contaminated with a model bacterium Rhodococcus erythropolis. The development of bacterial contamination can be expected during a real application of fungal bio filters in wastewater treatment. The main aim of the work was to study interspecies interactions in the model bioreactors during the dye decolorization. Ligninolytic enzyme activities were followed in the bioreactor cultures as markers of fungal biodegradation ability. In contrast to the controls, no bacterial growth was observed in the P. ostreatus bioreactor culture liquid. The results showed that fungal laccase, pH of the culture liquid, and glucose consumption by the fungus had no effect on the bacterial growth. However, 4*105 - 1,3*106 CFU/ml of R. erythropolis was detected to be associated with the fungal solid support. The presence of these bacteria had no effect on the decolorization performance of the bioreactors. Dye decolorization efficiency...
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The Occurence Of Bla(Tem) And Tet(W) Genes In Czech Wastewaters And Their Fate In Fungal-Based Wastewater Treatment Processes
Svobodová, Kateřina ; Adámek, M. ; Petráčková, Denisa
Antibiotic resistant bacteria and genes (ARB&Gs) have presently emerged as serious environmental pollutants with a high health risk. This study analyzed the occurrence of ARGs tet(W), bla(TEM) and intl1 in microbiomes from 4 Czech wastewater treatment plants showing that bla(TEM) displayed slightly higher gene abundance compared to tet(W). Next, simulated wastewater seeded with activated sludge bacteria was treated with Pleurotus ostreatus-based bioreactor to evaluate the effect of the fungal filter on bacterial community structure and bacterial resistome in wastewaters. The results showed that functional diversity of bacterial communities was significantly changed by their contact with fungal biofilm as revealed by EcoPlate assays and analyses of bacterial 16S rDNA by denaturing gradient gel electrophoresis. A significant decrease in tet(W) abundance (48 times) was observed in fungal-treated wastewater compared to untreated one. The abundance of bla(TEM) genes increased 4 times during the adaptation of bacteria to nutrient conditions and remained unaffected by the fungal treatment. To our knowledge this work is first to study the fate of ARGs in wastewaters during their advanced biological treatment. It documented that P. ostreatus has a potential to lower the abundance of some ARGs in wastewaters.
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Biodegradation of selected psychopharmaceuticals in underground water using Pleurotus ostreatus
Krejčová, Lucie ; Cajthaml, Tomáš (advisor) ; Pivokonský, Martin (referee)
The ability of the ligninolytic fungus Pleurotus ostreatusto degrade 4 pharmaceutical drugs and 5 compounds which are either used during drug manufacturing or are created as by-products was studied. These compounds were detected in groundwater near a drug manufacturing plant. The maximum concentration levels of the selected compounds in tested groundwater samples variedfrom0.23 µg/lto 227.87 µg/l apart from 1 compound which was not detected in any sample. The degradation efficiency of P. ostreatus was examined with individual compounds as well as with the mixture of all 9 compounds. When degrading individual compounds P. ostreatus lowered the initial concentration (10 mg/l) of 5 compounds by 62-100% after 14-day cultivation in malt extract-glucose medium. When degrading the compound mixture P. ostreatus lowered the initial concentration (2 mg/l of each compound) of 5 compounds by 50-100% after 14-day cultivation in malt extract-glucose medium. Acute toxicity tests with Vibrio fischeri suggest the formation of metabolites which are more toxic than the original compounds. The EC50 value for individual compounds during toxicity tests with Vibrio fischeri was 5.45-131.98 mg/l. Keywords:biodegradation, pharmaceuticals, ligninolytic fungi, Pleurotus ostreatus, groundwater, toxicity, Vibrio fischeri
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Biodegradation of polychlorinated biphenyls in underground water using biological approaches.
Šrédlová, Kamila ; Cajthaml, Tomáš (advisor) ; Innemanová, Petra (referee)
Polychlorinated biphenyls (PCBs) are a class of important organic pollutants which undergo very slow degradation and tend to persist for a long time in the environment. PCBs have various negative effects on living organisms, human health and the environment in general. A method for determining PCB levels in aqueous matrices was developed consisting of solid-phase extraction (SPE) and GC/MS determination. The average recoveries of PCB congeners from artificially spiked deionized water were around 85%. The PCB concentration of 3.3±0.1 μg·l-1 (a sum of selected congeners) was detected in contaminated groundwater originated from the area of a former asphalt concrete producing plant. The most abundant PCBs were trichlorinated congeners which together comprised about 80% of the total PCB content. The optimised SPE method was further applied together with accelerated solvent extraction (ASE) to assess the degradation potential of oyster mushroom (Pleurotus ostreatus) which shows high degradation efficiency of various organic pollutants including PCBs. P. ostreatus belongs to ligninolytic (white-rot) fungi, which is a very promising group of microorganisms frequently studied due to their bioremediation potential. P. ostreatus, strain KRYOS was able to remove 41±8% of the initial amount (50 μg) of the...
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Study of the interaction between fungus Pleurotus ostreatus and bacterial cultures on the abiotic surfaces - morphological, biochemical and proteomic analysis
Kozická, Barbora ; Petráčková, Denisa (advisor) ; Konopásek, Ivo (referee)
Ligninolytic fungi are well known for their ability to degrade a wide range of xenobiotics contaminating the environment, including synthetic industrial dyes. In this work Pleurotus ostreatus was used for decolorization of a synthetic textile dye Remazol Brilliant Blue R (RBBR). To set up a model fungal "fixed-bed" bioreactor the fungus was immobilized on a polyurethane foam and artificially contaminated with a model bacterium Rhodococcus erythropolis. The development of bacterial contamination can be expected during a real application of fungal bio filters in wastewater treatment. The main aim of the work was to study interspecies interactions in the model bioreactors during the dye decolorization. Ligninolytic enzyme activities were followed in the bioreactor cultures as markers of fungal biodegradation ability. In contrast to the controls, no bacterial growth was observed in the P. ostreatus bioreactor culture liquid. The results showed that fungal laccase, pH of the culture liquid, and glucose consumption by the fungus had no effect on the bacterial growth. However, 4*105 - 1,3*106 CFU/ml of R. erythropolis was detected to be associated with the fungal solid support. The presence of these bacteria had no effect on the decolorization performance of the bioreactors. Dye decolorization efficiency...
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Study of the metabolism of 17α-ethinylestradiol by cytochromes P450
Valášková, Petra ; Černá, Věra (advisor) ; Levová, Kateřina (referee)
A synthetic estrogen 17α-ethinylestradiol (EE2) is the main active component of the hormonal contraceptive pills. The rise of consumption of hormonal contraceptives has increased the risk of the back negative effects of EE2 to aquatic organisms. EE2 belongs to the endocrine disruptive compounds known for mimicking natural hormones. A more detailed examination of the transformation of this compound in vivo and in vitro can contribute to a better understanding of its negative effects. This master thesis is therefore devoted to the study of the metabolism of EE2 in two selected model organisms. The ligninolytic fungus Pleurotus ostreatus is the type of fungi with promising biodegradation ability to a lot of pollutants. These properties have led to numerous studies of the degradation potential of P. ostreatus towards EE2, with the possibility of removing this compound from the environment. EE2 has been degraded by the fungus P. ostreatus in vivo resulting in one hydroxylated metabolite, which estrogenic activity is in need for further study. In vitro studies were carried out with a microsomal fraction isolated from the mycelium of this fungus. The conversion of EE2 in vitro via CYPs dependent on NADPH has not been demonstrated, however using KHP as a cofactor, there was one metabolite of EE2 found,...
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Biodegradation of polychlorinated biphenyls by white - rot fungi and their enzymes
Linhartová, Lucie ; Bořek Dohalská, Lucie (advisor) ; Černá, Věra (referee)
Polychlorinated biphenyls (PCB) represent relevant persistent organopollutants of the environment and the estimated amount of PCB released into the environment is 750000 metric tons. White-rot fungi have been studied for long time due to their degradative potential toward various aromatic pollutants and it is known that these fungi are able to decompose PCB in vivo. Biodegradation of PCB by the fungus Pleurotus ostreatus was studied in the frame of this work. A high degradative efficiency of P. ostreatus was observed in the first set of experiments, even in the presence of relative high amount of added PCB. Fungus was able to transform 780±50 µg out of the intial amount 1000 µg in 20 ml of a cultivation media within 42 days. A decrease in toxicity was recorded during the degradation that suggests the suitability of this organism for a practical use in decontamination. In vitro experiments with purified laccase induced with Cu2+ from this fungus did not prove any participation of the enzyme in the first step of PCB transformation. The enzyme did not show an ability to degrade PCB even after purification from cultivation media containing PCB. It was found that the first step of PCB transformation can be performed by an intracellular process with microsomal fraction. A degradation of 44-67% was observed for...
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Substrates colonised by wood-inhabiting mycelia mushrooms and their technical utilization
Holejšovská, Eva ; Jablonský, Ivan (advisor) ; Súkeníková, Tereza (referee)
Cultivation of edible and medicinal mushrooms become of increasing importance worldwide. Fungi used as foods but also assist in human and veterinary medicine. The aim of this thesis was to prove that mushrooms may have technical applications. It was founded several attempts. Attempts have been founded on a substrate of straw pellets. The pellets had to be thoroughly pulverize the presence of water, at the most granular, in order to equalize the penetration of mushroom mycelia. Were observed following factors: growth rate, mycelium strength and thermal conductivity. The first two experiments were focused on the penetration rate, where it was found that mycelium grows fastest Lingzhi mushroom (Ganoderma lucidum), and Trametes versicolor (Trametes versicolor). One attempt was aimed at creating a bottle holder and lamp shades. At this experiment was used oyster mushroom (Pleurotus ostreatus). It was found that oyster with sufficient ingrowth without contamination creates strong mycelium can be used as packaging material. The theoretical part will learn about growing mushrooms selected, their appearance, and their medicinal effects of other cultivated species. Researched mushrooms were oyster mushroom (Pleurotus ostreatus), Lingzhi mushroom (Ganoderma lucidum), Ohňovec pine (Phellinus pini), Trametes versicolor (Trametes versicolor) and Lentinula edodes.
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