The primary purpose of this investigation is not only to improve the present technique of culture of Mycobacterium tuberculosis, but also to make a comparative study of the media use, namely, Veterans Administration modification of Trudeau's medium, Lowenstein's egg medium, and that developed by the author, in order to find which, if any, of these will most easily and effectively meet the needs of the hospital laboratory.

The main goal of this thesis is to expertise biological treatment of sewage water rising from production of DPG (diphenylguanidine). The theoretical part is focused on the water and means of sewage disposal. There is shortly described mechanism of physical, chemical, biological and combinated methods of sewage disposal. In practical part of this thesis, focused on expertising degradation of DPG and aniline was chosen high performance liquid chromatography (HPLC) technique, which made it possible to determined their biodegradation abilities. The next component of this thesis are sideline products determination, phenylurea especially. This thesis is focused on monitoring of biofilm´s character and the activity of Rhodococcus erythropolis bacterial culture, used in biodegradation of DPG in chemical-biological wastewater treatment plant, LZD a.s.

Since the beginning of the 20th century plastic has been a widely used material, which has resulted in large quantities of plastic being produced in the last century. The plastics of today are mainly produced from fossil raw materials, which gives it a high climate impact. Plastic also has a long service life, which creates problems with handling after the new period when new plastic is produced at a faster rate than plastic debris can be recycled or incinerated. One possible approach is to switch from plastic from fossil sources to bioplastics, which is produced by renewable sources. This means a reduction in the environmental impact as the amount of fossil CO2 emissions from combustion of plastics would decrease. Polyhydroxyalkanoates (PHAs) are created by short volatile fatty acids (VFAs) added to bio sludge from a wastewater treatment plant at a pulp and paper mill where there is a lot of bacteria and microorganisms. Some of the bacteria in the sludge have the ability to accumulate PHA when VFA is added in excess and then be able to use it as an energy and carbon source in cases of starvation. This means that PHA produced in this way can be degraded by bacteria making it biodegradable while having similar properties as oil-based plastics. Production of PHA is currently expensive as it is often necessary to purchase VFA for production. To make it economically sustainable to replace oil-based plastics with PHA, the cost of PHA production must therefore be reduced. This can be done by using mixed bacterial cultures from, for example, industrial wastewater treatment plants and by creating their own composition of VFA through fermentation. At the paper mill at BillerudKorsnäs Gruvön there are several process streams that today are led to the water treatment plant, one of which is from PM6 (Paper Machine 6). By fermenting that stream, VFA can be formed with a composition of acetic acid, propionic acid, and butyric acid. The current may thus be suitable to use as a substrate in PHA production. Another way to produce VFA is to ferment the residual flow from hydrothermal carbonation (HTC) of bio sludge. According to (Samorì et al., 2019), acetic acid, butyric acid and valeric acid are formed, which means that even this stream may be suitable as a substrate for PHA production. The purpose of this thesis is to investigate the effects of the composition of VFA on the production of PHA from forest industrial bio sludge. The study covers two different cases, Case PM6 and Case HTC, where two different types of composition of VFA are added to paper sludge from the paper mill. The experiments were performed in cylindrical tanks on three occasions where the first two experiments had a volume of 30 liters and the last experiment a volume of 10 liters. On the first occasion, the maximum accumulation rate in the growth phase was studied. In the second instance, a high accumulation rate was sought, but also survival after the growth phase. The first two trials were batch trials where the sludge was dosed until saturation was achieved. On the third occasion, the possibility of carrying out PHA production with a continuous sludge exchange was studied. The experiments were analyzed by FTIR which provided information on the absorbance of the sludge which shows how the PHA concentration increased during the course of the experiments. Extractions were then performed to obtain the concentration of PHA that eventually accumulated in the sludge. The results show that biomass from BillerudKorsnäs Gruvön's mills accumulated PHA faster with VFA composition from fermented PM6 effluent compared to VFA composition from fermented HTC condensate. Calculations made with input from the experiments indicate that it is possible to produce a larger amount of PHA per year with Case PM6. The conclusion is therefore that Case PM6 is preferable if as large a PHA production as possible wants to be achieved. Case HTC is instead preferred if reduced PHA production can be tolerated in favor of biocarbon production.

Submitted by Deyse Queiroz (deysequeirozz@hotmail.com) on 2018-05-22T15:00:44Z No. of bitstreams: 1 JESSÉ MALVEIRA GOMES - DISSERTAÇÃO PPGHT - 2017.pdf: 1522466 bytes, checksum: cc3a2a2cb3b9b312f031097d4500a8df (MD5) / Made available in DSpace on 2018-05-22T15:00:44Z (GMT). No. of bitstreams: 1 JESSÉ MALVEIRA GOMES - DISSERTAÇÃO PPGHT - 2017.pdf: 1522466 bytes, checksum: cc3a2a2cb3b9b312f031097d4500a8df (MD5) Previous issue date: 2017-03 / O milho é uma das culturas mais produzida e importante no mundo, sendo utilizada na alimentação humana e animal. O Brasil é um dos maiores produtores desse grão, sendo o terceiro maior produtor do mundo. Apesar de uma produção elevada, a produtividade é comprometida pela ação de insetos pragas, sendo a Spodoptera frugiperda Smith a de maior atuação nessa cultura. Em meio a esse desafio, o uso constante de inseticidas é utilizado com intuito de minimizar as perdas nas lavouras, porém, ocasionando grandes impactos ambientais, bem como a contaminação de trabalhadores rurais. Uma das estratégias viáveis e promissoras na supressão de pragas tem sido o manejo integrado de pragas (MIP), que tem como alicerce o uso isolado ou consorciado de técnicas de controles de pragas, sendo o controle biológico uma das ferramentas mais sustentáveis do ponto de vista ecológico. Desta forma, o presente trabalho teve como objetivo avaliar a toxicidade da cepa bacteriana 358.1 isolada do petróleo e o sobrenadante da cultura de células sobre o controle de lagartas de 2º e 3º instares de S. frugiperda, comparando as taxas de mortalidade e efeitos sobre seu desenvolvimento e com inseticidas químicos comerciais. O experimento consistiu de duas etapas, o primeiro ensaio foi realizado com lagartas de 3º instar com três diluições diferentes da cepa 358.1, sobrenadante da cultura de células, solução salina como controle negativo e o inseticida fenpropatrina como controle positivo. No segundo ensaio utilizou-se lagartas de 3º e 2º instares, a cultura celular da cepa 358.1 em sua concentração total (4 x 108 UFC/mL), seu sobrenadante da cultura, como controle negativo água destilada e testemunha positiva o produto comercial flubendiamida. Os experimentos com a cepa 358.1, sobrenadante da cultura de células e os produtos químicos testados não resultaram em uma taxa de mortalidade de S. frugiperda significante, porém, a cepa 358.1 e seu sobrenadante da cultura resultaram no aumento da duração da fase larval e em distúrbios morfofisiológicos do inseto, ocasionando, inclusive, no surgimento de adultos com asas atrofiadas, sendo desta maneira, promissores para à realização de novos testes. / Corn is one of the most produced and important crops in the world, utilized for human and animal feeding. Brazil is one of the top producers of this grain, being the third biggest producer worldwide. Despite the great production, the productivity is compromised due to the activity of insect pest. Because of this challenge, the continuous use of insecticides is done aiming to minimize lost in the crops. However, it causes major environmental impact and it is also contaminating rural workers. One of the viable and promising strategies to the pest suppression has been the pest integrated management (PIM), which is based on the use of isolated or consortium of pest controlling techniques, the biological control being one of the most sustainable tools in an ecological perspective. Therefore, the present study had the goal of evaluating the toxicity of the 358.1 bacterium strain isolated from the oil and supernatant of the cell culture under control of caterpillars from 2nd and 3rd instars of S. frugiperda, comparing the death rates and effects on their development, and also comparing with commercial chemical insecticides. The experiment consisted in two stages; the first assay was done with 3rd instar caterpillar with three different dilutions from the 358.1 strain, supernatant of the cell culture, saline solution as negative control, and fenpropatrina insecticide as positive control. On the second assay, 3rd and 2nd instars caterpillars were used, the cell culture from the 358.1 strain on its total concentration (4 x 108 UFC/mL), supernatant of the cell culture, distilled water as negative control and flubendiamida, a commercialized product, as the positive control. The experiments with the 358.1 strain, supernatant of the cell culture and the tested chemical products didn’t show a significant death rate for S. frugiperda, however, the 358.1 strain and supernatant of the cell culture resulted in an increase in the larval phase duration and on morphophysiological disturbs on the insect, resulting yet in the emergence of adults with atrophied wings, thus being promising to the realization of new tests.

Chlorbenzole sind aufgrund ihrer weiten Verbreitung in Industrie und Landwirtschaft und ihrer geringen chemischen Reaktivität sowie guten Fettlöslichkeit ubiquitäre Umweltkontaminanten, die sich in der Nahrungskette anreichern. Eine natürliche, mikrobielle Dechlorierung dieser Verbindungen ist von besonderem Interesse, da die Toxizität chlorierter Benzole mit der Anzahl der Chlorsubstituenten steigt. Im Gegensatz zu Mono- und Dichlorbenzol, die durch aerobe Laborreinkulturen dechlorierbar sind, werden höher chlorierte Benzole bevorzugt unter anaeroben Bedingungen in Mischkulturen mit unbekannter Spezieszusammensetzung dechloriert. Bioreaktoren, in denen solche undefinierten Mischkulturen kontinuierlich kultiviert werden, sind eine vielversprechende Technik bei der Behandlung Chlorbenzol-kontaminierter Abwässer. Aufgrund der unbekannten Zusammensetzung der Population muß die mikrobielle Aktivität jedoch als "black box" betrachtet werden, was eine direkte Steuerung und Optimierung solcher Bioreaktoren erschwert. Zur Bestimmung der mikrobiellen Diversität einer in ihrer Zusammensetzung unbekannten, Trichlorbenzol(TCB)-dechlorierenden Bioreaktorkultur wurde aufgrund der bekannten Limitierungen kulturabhängiger Methoden die kulturunabhängige, vergleichende Sequenzanalyse direkt amplifizierter und klonierter 16S-rDNA gewählt. Die durch eine neuentwickelte Hybridisierungsmethode wesentlich vereinfachte Analyse der 16S-rDNA-Genbibliotheken erlaubte eine phylogenetische Klassifizierung der in der TCB-dechlorierenden Mischkultur abundanten Mikroorganismen (Bakterien und Archaea) und zeigte das Auftreten von 51 bakteriellen 16S-rDNA-Klonfamilien, mit einem weiten phylogenetischen Spektrum und teilweise enge Verwandtschaften zu anaerob dechlorierenden Dehalobacter spp. oder zu unkultivierten Bakterien eines vergleichbaren Biotops. Dieser Diversität stand eine dominierende Methanosaeta-ähnliche Klonfamilie innerhalb der Archaea gegenüber. Die aus der phylogenetischen Klassifizierung abgeleiteten metabolischen Eigenschaften einiger Bakterien und Archaea der TCB-dechlorierenden Mischkultur konnten durch gezielte Anreicherung und in-vitro Kultivierung bzw. die kulturunabhängige Sequenzanalyse funktioneller Gene bestätigt werden. Die dargestellten Ergebnisse lassen eine spezifische Zusammensetzung der TCB-dechlorierenden Mischkultur vermuten und geben Hinweise auf Indikatororganismen, die ein Monitoring und eine damit verbundene Effizienzsteigerung der anaeroben TCB-Dechlorierung im Bioreaktor ermöglichen könnten. / Due to their widespread application in industry and agriculture and their chemical stability chlorobenzenes (CB) are ubiquitous pollutants in soil, sediments, and aquifers. Since toxicity of CBs increases with the number of chlorine substituents, microbial dechlorination of CBs is of major interest. In contrast to di- and monochlorobenzenes (DCB and MCB) higher chlorinated benzenes are more resistant to aerobic dechlorination. However, for these compounds reductive dechlorination by different anaerobic microbial communities is well known. Bioreactors inoculated with complex dechlorinating anaerobic microbiota seem to be promising technologies for bioremediation of CB-contaminated aquifers. Several studies showed the efficiency of such bioreactors in treating chloroaromatic contaminated wastewaters. However, due to the unknown species diversity microbial activity had to be treated as a "black box" and direct optimization was hampered. To determine the microbial diversity of an anaerobic consortium in a fluidized bed reactor used for dechlorination of trichlorobenzene (TCB) I employed comparative sequence analysis of 16S rRNA genes after direct PCR-amplification and cloning from community DNA since culture-based methods have shown to be strongly biased. The application of a new hybridization based screening approach for bacterial 16S rDNA clone libraries drastically simplified the analysis and allowed the phylogenetic classification of the abundant bacteria and archaea. A total of 51 bacterial 16S rDNA clone families were found, which showed a wide distribution among the main bacterial phyla. Several bacterial 16S rDNA clone families were closely related to anaerobic, dechlorinating Dehalobacter spp. and to yet-uncultured bacteria of a similar habitat. In contrast to the high bacterial diversity archaeal 16S rDNA clone libraries were clearly dominated by a Methanosaeta concilii-like clone family. Some yet-uncultured bacteria and archaea of the TCB-dechlorinating consortium were sufficiently closely related to studied organsims that reasonable physiological hypotheses could be formulated. These hypotheses were confirmed by either cultivation of the respective organism or by culture-independent sequence analysis of specific functional genes. The results suggest a specific community structure of the TCB-dechlorinating consortium and give evidence for indiator organisms. Moleculargenetic monitoring of these indicator organisms might allow the optimization of the continous TCB-dechlorination in the fluidized bed reactor.