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Biotechnological production of PHA by selected bacterial isolates
Matějka, Filip ; Šedrlová, Zuzana (referee) ; Obruča, Stanislav (advisor)
This bachelor thesis deals with the production of polyhydroxyalkanoates (PHA) using thermophilic bacterial isolates with designation 34, 35 and BŽ. Bacterial straines were isolated from activated sludge and compost The theoretical part contains a description of PHA, extremophilic bacteria and detection techniques for the determination of polyhydroxyalkanoates. In the experimental part, the presence of the phaC gene, which is crucial for the ability to produce PHA, was first determined by PCR and bacterial strains were also subjected to DNA sequencing of 16S rRNA gene which enabled preliminary taxonomical classification of the isolates. In the next part of the experimental work, the conditions for biomass growth and PHA production were optimized. Suitable carbon source, the ideal temperature for cultivation and the influence of precursors on the production of copolymers were studied and identified. The composition and proportion of PHA were determined spectrophotometrically and by GC-FID. Finally, visual screening of PHA accumulation inside bacterial cells was performed using fluorescence microscopy.
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Microfauna of the activated silt from the biological sewage disposal plant in Kroměříž region.
Hrudíková, Radka ; Illková, Kateřina (referee) ; Omelková, Jiřina (advisor)
Bachelor thesis deals with the description and evaluation of micro fauna of activated sludge biological sewage waste water in Kromeriz region. Furthermore, morphological description and evaluation of sludge flocs. The thesis describes the types of biota (biocenosis), which may occur in the activation tanks. There are also mentioned other incidental methods of sewage treatment such as mechanical, chemical and physicochemical methods. The experimental section describes the experiment procedure, apparatus and methods use for assessment of activated sludge. In the results the data are processed into the synoptic charts and the accompanying tables. At the end the results are evaluated.
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Monitoring of selected parameters in the wastewater treatment process in the various WWTP
Dalajková, Nikola ; Zlámalová Gargošová, Helena (referee) ; Repková, Martina (advisor)
This work is focused on wastewater treatment, specifically the WWTP Brno Modřice. Described herein are the general technological parameters of large and medium-sized wastewater treatment plants and wastewater treatment process in the WWTP Brno Modřice. The greatest emphasis is on biological phase of waste water treatment. Purification employs chemical, biological and physical cleaning processes. To determine the effectivness of the treatment plants various indicators are used such as COD, BOD, Ntotal and Ptotal whose the maximum permissible values are established in accordance to norm. The basis of biological treatment is the activated sludge containing microorganisms, by which the effectiveness of the cleaning process is determined. These microorganisms are determined qualitatively and quantitatively. To compare the efficiency measurements of activated sludge were also made at two other wastewater treatment plants. These values were then compared in the experimental part of the work.
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Level scraping systems of settlement tanks
Bartoš, Pavel ; Medlík, Jan (referee) ; Brandejs, Jan (advisor)
This bachelor’s thesis contains survey of various ways to clean waste water, types of waste treatment plants and its components. The thesis above all adverts to two types of waste treatment tank bottom raking. It describes individual raking effects, advantages, disadvantages and convenience of use. Closing disserts on methods of sludge thickening with use of thickening tanks.
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Upgrading of wastewater treatment plant by means of MBR technology.
Spratková, Aneta ; Rusník,, Igor (referee) ; Hlavínek, Petr (advisor)
This diploma thesis is focused on the topic of membrane technologies used in wastewater treatment. The research part of the thesis deals with membrane separation of activated sludge, and the focus is on the submerged membrane modules. The thesis contains basic principles of MBR systems, an overview of used membrane modules, modes of operation, advantages and disadvantages of using this process. The practical part of the thesis proposes the intensification of WWTP Perná with using MBR technology. This part includes the technical-economic assessment of the activation WWTP with the third stage of treatment and activation WWTP with MBR.
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Isolation, identification and characterization of extremophiles capable of PHA production
Vlasáková, Terézia ; Turková, Kristýna (referee) ; Obruča, Stanislav (advisor)
This diploma thesis is focused on isolation and identification of thermophilic microorganisms capable of production of polyhydroxyalkanoates (PHA) in the sample of activated sludge from wastewater treatment. 6 culture samples were isolated from activated sludge by means of cultivation technics and methods of molecular biology. They were closer specified by comparing nucleotide sequences of 16S-rRNA gene and assigned to bacterial genus Anoxybacillus. The production of PHA by this genus was not reported in literature so far. Samples were confirmed to contain phaC gene that codes the enzyme PHA-synthase and they also gave a positive response to staining colonies with Nile red, what refers to presence of intracellular lipidic structures. However, the PHA production by isolates was not successful. The reason should be an inappropriate production medium or conditions. The positive phenotype result of Nile red dyeing was probably achieved by production of huge amount of lipids by bacterial cells that provides similar fluorescence than PHA granules.
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Isolation of PHA producing bacteria from mixed microbial consortia
Plachý, Petr ; Kučera, Dan (referee) ; Obruča, Stanislav (advisor)
Aim of this bachelor thesis is detection of polyhydroxyalcanoates (PHA) producing bacteria from activated sludge and effort for isolation of these bacteria. The theoretical part deals with general issues of PHA and of bacterial production of PHA. Also there is attention paid to characterization of activated sludge and to selected methods used in this thesis for detection of PHA producing bacteria. In the experimental part, mixed microbial culture of the activated sludge was cultivated on different carbon sources. Potential PHA producers was isolated from these cultures with the use of lipophilic staining with Nile red and phaC gene (essential for PHA synthesis) was detected with the use of polymerase chain reaction (PCR) in 11 of the isolated cultures. By DNA sequencing 8 bacterial cultures were identified. It was Klebsiella pneumoniae (1 x), Paenirhodobacter enshiensis (1 x) and Pseudomonas putida (6 x). Presence of PHA in biomass was detected in 2 of the 11 isolated cultures by Fourier transformation infrared spectroscopy (FTIR) analysis. The content of polyhydroxybutyrate (PHB) was determined with the use of gas chromatography to 9,33 % of dry biomass (Paenirhodobacter enshiensis) and to 1,18 % (unidentified culture).
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Activated sludge inhibited by pesticides
Pokluda, Michal ; Pavlíková, Marcela (referee) ; Hrich, Karel (advisor)
The bachelor's thesis focuses on researching the inhibitory effects of selected pesticide substances on activated sludge present in wastewater treatment plants. The thesis is divided into a theoretical part and a practical part. The first part provides a theoretical explanation of pesticides, their presence in water, their basic classification, and the legislation addressing the issue of pesticide substances in water. The thesis also lists and describes methods for removing pesticides from water. Furthermore, the content of the theoretical part touches upon the topic of the Modřice wastewater treatment plant, primarily describing its treatment process. The practical part of the thesis involves laboratory research on the oxygen consumption rate of microorganisms in activated sludge in the presence of pesticides toxic to these organisms. The testing methodology is based on the standard ČSN EN ISO 8192.
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Biotechnological production of PHA by selected bacterial isolates
Matějka, Filip ; Šedrlová, Zuzana (referee) ; Obruča, Stanislav (advisor)
This bachelor thesis deals with the production of polyhydroxyalkanoates (PHA) using thermophilic bacterial isolates with designation 34, 35 and BŽ. Bacterial straines were isolated from activated sludge and compost The theoretical part contains a description of PHA, extremophilic bacteria and detection techniques for the determination of polyhydroxyalkanoates. In the experimental part, the presence of the phaC gene, which is crucial for the ability to produce PHA, was first determined by PCR and bacterial strains were also subjected to DNA sequencing of 16S rRNA gene which enabled preliminary taxonomical classification of the isolates. In the next part of the experimental work, the conditions for biomass growth and PHA production were optimized. Suitable carbon source, the ideal temperature for cultivation and the influence of precursors on the production of copolymers were studied and identified. The composition and proportion of PHA were determined spectrophotometrically and by GC-FID. Finally, visual screening of PHA accumulation inside bacterial cells was performed using fluorescence microscopy.
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Isolation of PHA producing bacteria from mixed microbial consortia
Plachý, Petr ; Kučera, Dan (referee) ; Obruča, Stanislav (advisor)
Aim of this bachelor thesis is detection of polyhydroxyalcanoates (PHA) producing bacteria from activated sludge and effort for isolation of these bacteria. The theoretical part deals with general issues of PHA and of bacterial production of PHA. Also there is attention paid to characterization of activated sludge and to selected methods used in this thesis for detection of PHA producing bacteria. In the experimental part, mixed microbial culture of the activated sludge was cultivated on different carbon sources. Potential PHA producers was isolated from these cultures with the use of lipophilic staining with Nile red and phaC gene (essential for PHA synthesis) was detected with the use of polymerase chain reaction (PCR) in 11 of the isolated cultures. By DNA sequencing 8 bacterial cultures were identified. It was Klebsiella pneumoniae (1 x), Paenirhodobacter enshiensis (1 x) and Pseudomonas putida (6 x). Presence of PHA in biomass was detected in 2 of the 11 isolated cultures by Fourier transformation infrared spectroscopy (FTIR) analysis. The content of polyhydroxybutyrate (PHB) was determined with the use of gas chromatography to 9,33 % of dry biomass (Paenirhodobacter enshiensis) and to 1,18 % (unidentified culture).
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