Dominant genera of bacteria were isolated from three river waters during anthracene and pentachlorophenol biotransformation studies. The genera Pseudomonas, Acinetobacter, Micrococcus, Chromobacterium, Alcaligenes, Azomonos, Bacillus, and Flavobacterium were capable of biotransforming one or both of these compounds. These isolates were subjected to further biotransformation tests, including river water and a basal salt medium with and without additional glucose. The results of these experiments were evaluated statistically. It was concluded that only a limited number of the bacteria identified were able to transform these chemicals in river water. The addition of glucose to the growth medium significantly affected the biotransformation of these chemicals. It was also determined that the size of the initial bacterial population is not a factor in determining whether biotransformation of anthracene or pentachlorophenol can occur.

by Law Wing Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 192-206). / Abstracts in English and Chinese. / Acknowledgments --- p.i / Abstract --- p.ii / List of Figures --- p.vi / List of Tables --- p.xii / Abbreviations --- p.xv / Chapter 1. --- Introduction / Chapter 1.1. --- Pesticides --- p.1 / Chapter 1.1.1. --- Types and uses --- p.1 / Chapter 1.1.2. --- Development of pesticides --- p.1 / Chapter 1.1.3. --- The case against pesticides --- p.3 / Chapter 1.2. --- Pentachlorophenol --- p.4 / Chapter 1.2.1. --- Production --- p.4 / Chapter 1.2.2. --- Toxicity --- p.4 / Chapter 1.2.3. --- Persistency --- p.6 / Chapter 1.3. --- Methyl-parathion --- p.9 / Chapter 1.3.1. --- Production --- p.9 / Chapter 1.3.2. --- Toxicity --- p.9 / Chapter 1.3.3. --- Environmental fate --- p.12 / Chapter 1.4. --- Conventional methods dealing with pesticides --- p.12 / Chapter 1.5. --- Bioremediation --- p.15 / Chapter 1.6. --- Spent mushroom compost --- p.17 / Chapter 1.6.1. --- Background --- p.17 / Chapter 1.6.2. --- "Physical, chemical and biological properties of SMC " --- p.19 / Chapter 1.6.3. --- Recycling of agricultural residuals --- p.21 / Chapter 1.6.3.1. --- Definition --- p.21 / Chapter 1.6.3.2. --- Types of recycling --- p.22 / Chapter 1.6.4. --- Potential uses of SMC as bioremediating agent --- p.23 / Chapter 1.6.4.1. --- Use of microorganisms in SMC --- p.23 / Chapter 1.6.4.2. --- Use of ligninolytic enzymes in SMC --- p.24 / Chapter 1.7. --- Ligninolytic enzymes --- p.28 / Chapter 1.7.1. --- Background --- p.28 / Chapter 1.7.2. --- What are white rot fungi? --- p.29 / Chapter 1.7.3. --- Why is lignin so difficult to degrade? --- p.29 / Chapter 1.7.4. --- Three main ligninolytic enzymes --- p.32 / Chapter 1.7.4.1. --- Lignin peroxidases (LiP) --- p.32 / Chapter 1.7.4.2. --- Manganese peroxidase (MnP) --- p.36 / Chapter 1.7.4.3. --- Laccase --- p.37 / Chapter 1.8. --- Why SMC was chosen to be the bioremediating agent in my project? --- p.40 / Chapter 1.9. --- Bioremediation of chlorophenols and PCP --- p.44 / Chapter 1.9.1. --- Bacterial system --- p.44 / Chapter 1.9.2. --- Fungal system --- p.45 / Chapter 1.10. --- Bioremediation of methyl-parathion --- p.49 / Chapter 1.10.1. --- Bacterial system --- p.49 / Chapter 1.10.2. --- Fungal system --- p.51 / Chapter 1.11. --- Proposal and experimental plan of the project --- p.51 / Chapter 1.11.1. --- Study the removal of pesticides in both aquatic and soil system --- p.52 / Chapter 1.11.2. --- Research strategy --- p.52 / Chapter 1.11.3. --- Optimization of pesticide removal --- p.53 / Chapter 1.11.4. --- Identification of breakdown products --- p.54 / Chapter 1.11.5. --- Toxicity assay --- p.54 / Chapter 1.11.6. --- Isotherm plot --- p.55 / Chapter 1.12. --- Objectives of the study --- p.56 / Chapter 2. --- Material and Methods --- p.58 / Chapter 2.1. --- Material --- p.59 / Chapter 2.2. --- Production of Spent Mushroom Compost (SMC) --- p.59 / Chapter 2.3. --- Characterization of SMC --- p.60 / Chapter 2.3.1. --- PH --- p.60 / Chapter 2.3.2. --- Electrical conductivity --- p.60 / Chapter 2.3.3. --- "Carbon, hydrogen, nitrogen and sulphur contents " --- p.60 / Chapter 2.3.4. --- Ash content --- p.61 / Chapter 2.3.5. --- Metal analysis --- p.61 / Chapter 2.3.6. --- Anion content --- p.62 / Chapter 2.3.7. --- Chitin assay --- p.62 / Chapter 2.4. --- Characterization of soil --- p.63 / Chapter 2.4.1. --- Soil texture --- p.63 / Chapter 2.4.2. --- Moisture content --- p.64 / Chapter 2.5. --- Basic studies on the removal capacity of pesticides by SMC --- p.65 / Chapter 2.5.1. --- Preparation of pentachlorophenol and methyl- parathion stock solution --- p.66 / Chapter 2.6. --- Experimental design --- p.65 / Chapter 2.6.1. --- In aquatic system --- p.65 / Chapter 2.6.2. --- In soil system --- p.68 / Chapter 2.7. --- Extraction of pesticides --- p.68 / Chapter 2.7.1. --- In aquatic system --- p.68 / Chapter 2.7.2. --- In soil system --- p.69 / Chapter 2.8. --- Quantification of pesticides --- p.69 / Chapter 2.8.1. --- By high performance liquid chromatography --- p.69 / Chapter 2.8.2. --- By gas chromatography-mass spectrometry --- p.71 / Chapter 2.9. --- Optimization of pesticides degradation by SMC in both aquatic and soil systems --- p.72 / Chapter 2.9.1. --- Effect of initial pesticide concentrations on the removal of pesticides --- p.72 / Chapter 2.9.2. --- Effect of amount of SMC used on the removal of pesticides --- p.73 / Chapter 2.9.3. --- Effect of incubatoin time on the removal of pesticides --- p.73 / Chapter 2.9.4. --- Effect of initial pH on the removal of pesticides --- p.73 / Chapter 2.9.5. --- Effect of incubation of temperature on the removal of pesticides --- p.74 / Chapter 2.10. --- The study of breakdown process of pesticides --- p.74 / Chapter 2.10.1. --- GC/MS --- p.74 / Chapter 2.10.2. --- Ion chmatography --- p.74 / Chapter 2.11. --- Microtox® assay --- p.75 / Chapter 2.12. --- Assessment criteria --- p.75 / Chapter 2.12.1. --- In aquatic system --- p.75 / Chapter 2.12.2. --- In soil system --- p.76 / Chapter 2.13. --- Statistical analysis --- p.77 / Chapter 3. --- Results / Chapter 3.1. --- Characterization of SMC and soil --- p.78 / Chapter 3.2. --- Quantification of pesticides by HPLC and GC/MS --- p.82 / Chapter 3.3. --- Extraction efficiencies of pesticides with hexane --- p.82 / Chapter 3.4. --- Stability of pesticides against time --- p.82 / Chapter 3.5. --- Effect of sterilization of soil in the removal abilities of pesticides…… --- p.88 / Chapter 3.6. --- Optimization of removal of pentachlorophnol --- p.88 / Chapter 3.6.1. --- Effect of incubation time --- p.88 / Chapter 3.6.1.1. --- In aquatic system --- p.88 / Chapter 3.6.1.2. --- In soil system --- p.88 / Chapter 3.6.2. --- Effect of initial PCP concentrations and amout of SMC used --- p.91 / Chapter 3.6.2.1. --- In aquatic system --- p.91 / Chapter 3.6.2.2. --- In soil system --- p.94 / Chapter 3.6.3. --- Effect of pH --- p.97 / Chapter 3.6.3.1. --- In aquatic system --- p.97 / Chapter 3.6.3.2. --- In soil system --- p.97 / Chapter 3.6.4. --- Effect of incubation temperature --- p.97 / Chapter 3.6.4.1. --- In aquatic system --- p.97 / Chapter 3.6.4.2. --- In soil system --- p.101 / Chapter 3.6.5. --- Potential breakdown intermediates and products --- p.101 / Chapter 3.6.5.1. --- In aquatic system --- p.101 / Chapter 3.6.5.2. --- In soil system --- p.104 / Chapter 3.7. --- Microtox® assay of PCP --- p.110 / Chapter 3.7.1. --- In aquatic system --- p.110 / Chapter 3.7.2. --- In soil system --- p.110 / Chapter 3.8. --- Optimization of removal of methyl-parathion --- p.113 / Chapter 3.8.1. --- Effect of incubation time --- p.113 / Chapter 3.8.1.1. --- In aquatic system --- p.113 / Chapter 3.8.1.2. --- In soil system --- p.113 / Chapter 3.8.2. --- Effect of initial concentration and amount of SMC --- p.115 / Chapter 3.8.2.1. --- In aquatic system --- p.115 / Chapter 3.8.2.2. --- In soil system --- p.117 / Chapter 3.8.3. --- Effect of incubation temperature --- p.120 / Chapter 3.8.3.1. --- In aquatic system --- p.120 / Chapter 3.8.3.2. --- In soil system --- p.120 / Chapter 3.8.4. --- Potential breakdown intermediates and products --- p.121 / Chapter 3.8.4.1. --- In aquatic system --- p.121 / Chapter 3.8.4.2. --- In soil system --- p.124 / Chapter 3.9. --- Microtox ® assay of methyl-parathion --- p.133 / Chapter 3.9.1. --- In aquatic system --- p.133 / Chapter 3.9.2. --- In soil system --- p.133 / Chapter 4. --- Discussion / Chapter 4.1. --- Characterization of SMC and soil --- p.137 / Chapter 4.2. --- Stability of pesticides against time in aquatic and soil system --- p.141 / Chapter 4.3. --- Effect of sterilization of soil in the removal abilities of pesticides --- p.142 / Chapter 4.4. --- Optimization of removal of PCP --- p.142 / Chapter 4.4.1. --- Effect of incubation time --- p.142 / Chapter 4.4.1.1. --- In aquatic system --- p.142 / Chapter 4.4.1.2. --- In soil system --- p.143 / Chapter 4.4.2. --- Effect of initial PCP concentrations and amount of SMC --- p.144 / Chapter 4.4.2.1. --- In aquatic system --- p.144 / Chapter 4.4.2.2. --- In soil system --- p.147 / Chapter 4.4.3. --- Effect of pH --- p.149 / Chapter 4.4.3.1. --- In aquatic system --- p.149 / Chapter 4.4.3.2. --- In soil system --- p.150 / Chapter 4.4.4. --- Effect of incubation temperature --- p.150 / Chapter 4.4.4.1. --- In aquatic system --- p.150 / Chapter 4.4.4.2. --- In soil system --- p.152 / Chapter 4.4.5. --- Potential breakdown intermediates and products --- p.152 / Chapter 4.4.5.1. --- In aquatic system --- p.152 / Chapter 4.4.5.2. --- In soil system --- p.158 / Chapter 4.5. --- Microtox® assay of PCP --- p.159 / Chapter 4.5.1. --- In aquatic system --- p.159 / Chapter 4.5.2. --- In soil system --- p.160 / Chapter 4.6. --- Removal of PCP by the aqueous extract of SMC --- p.162 / Chapter 4.7. --- Optimization of removal of methyl-parathion --- p.164 / Chapter 4.7.1. --- Effect of incubation time --- p.164 / Chapter 4.7.1.1. --- In aquatic system --- p.164 / Chapter 4.7.1.2. --- In soil system --- p.165 / Chapter 4.7.2. --- Effect of initial methyl-paration concentrations and amount of SMC used --- p.165 / Chapter 4.7.2.1. --- In aquatic system --- p.165 / Chapter 4.7.2.2. --- I in soil system --- p.166 / Chapter 4.7.3. --- Effect of incubation temperature --- p.168 / Chapter 4.7.3.1. --- In aquatic system --- p.168 / Chapter 4.7.3.2. --- In soil system --- p.169 / Chapter 4.7.4. --- Potential breakdown intermediates and products --- p.169 / Chapter 4.7.4.1. --- In aquatic system --- p.169 / Chapter 4.7.4.2. --- In soil system --- p.170 / Chapter 4.8. --- Microtox® assay of Methyl-parathion --- p.173 / Chapter 4.8.1. --- In aquatic system --- p.173 / Chapter 4.8.2. --- In soil system --- p.174 / Chapter 4.9. --- Removal of methyl-parathion by the aqueous extract of SMC --- p.174 / Chapter 4.10. --- The ability of different types of SMC in the removal of organic pollutants --- p.176 / Chapter 4.11. --- The storage of SMC --- p.178 / Chapter 4.12. --- The effect of scale in the removal of pesticides --- p.180 / Chapter 4.13. --- Cost-effectiveness of using SMC as crude enzymes sources --- p.180 / Chapter 4.14. --- The effect of surfactant on the removal of PCP --- p.182 / Chapter 4.15. --- Prospects for employment SMC in removal of pollutants --- p.185 / Chapter 5. --- Conclusions --- p.186 / Chapter 6. --- Future investigation --- p.190 / Chapter 7. --- References --- p.192

Pentachlorophenol (PCP) is a biocide used by the NZ forestry industry until 1988. Its use was discontinued due to its toxicity to humans and animals. White-rot fungi have been shown to degrade PCP in laboratory and field trials. New Zealand native white-rot fungi were screened to identify organisms suitable for the clean up of PCP contaminated sites. Four criteria were used for the screening: fungal growth at different temperatures, PCP and creosote resistance and PCP degradation in standard liquid medium. Twenty isolates were identified as potentially useful from over 200 that were screened. One unique isolate, Trametes sp. HR577, was chosen for intensive study because it produced the well known laccases previously described from other PCP-degrading white-rot fungi. The white-rot isolate HR577 was assigned to the genus Trametes based on morphological characteristics and gene sequencing studies. The latter showed that the partial laccase gene sequences from Trametes sp. HR 577 had high sequence homology to laccases from other Trametes species, especially T. versicolor and T. villosa. Two laccase isozymes, designated L1c and L2, were purified from Trametes sp. HR577. These isozymes had similar biological properties to other Trametes species laccase isozymes. Both isozymes had a relatively high temperature optima, however, they were not very stable at elevated temperature. The dependence of laccase on dissolved oxygen for catalysis was demonstrated for isozyme L2. Laccase activity was severely inhibited in the absence of dissolved oxygen. This could be restored by reoxygenation into the assay system. Whole cultures of Trametes sp. HR577 grown in liquid culture removed up to 76% of PCP after 72 hours. PCP removal was mostly due to degradation rather than adsorption of PCP to fungal mycelium. Addition of purified and crude laccase isozymes (100 U mL-1) did not enhance PCP degradation. 6-15% of PCP was removed from solutions containing solely purified isozyme L1c or L2 in acetate buffer over 72 hours. Addition of ethanol or the laccase mediator compound 2,2' azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) resulted in increased PCP disappearance from purified laccase cultures. These studies show that the white-rot Trametes sp. HR577 has potential to be used for the clean up of PCP contaminated sites in NZ.

Chlorinated aromatic compounds are an important group of compounds. Many of them have been produced in large quantities and they are indispensable to technological and societal benefits. But regulatory agencies have tightened regulations on the use and release of chlorinated aromatic compounds because of the scientific understanding of their toxicity, persistence, behavior in the environment and their potential to cause adverse effects on the ecosystem and human health. / Pentachlorophenol (PCP), octachloronaphthalene and decachlorobiphenyl are all highly chlorinated aromatic compounds, of which, PCP has been used mainly as a biocide. Octachloronaphthalene and decachlorobiphenyl don't have practical use, but their congeners have been used widely as chemicals in industry. These compounds are toxic, recalcitrant and bio-accumulated within organisms. As the conventional treatment, incineration of these compounds can cause more serious problems, so that suitable alternatives need to be developed for their detoxification. / When compared with biodegradation or the thermal treatment of these compounds, chemical degradations have several merits. (Abstract shortened by UMI.)

O solo é um dos compartimentos mais atingidos pelo acúmulo de poluentes de origem antrópica, pois atua como depósito de poluentes e como fonte para outros ambientes de interface. Este estudo investigou sítio contaminado com resíduos da indústria de preservativos de madeira como fonte de compostos mutagênicos; definiu rotas e abrangência na dispersão desses contaminantes pela remobilização de partículas e deposição atmosférica, associando riscos ambientais e à saúde. Foram selecionados locais de amostragem para solos e poeira domiciliar a distâncias gradativas a partir de SI (pool de solo da área industrial), SR150 (150m), SR500 e SR1700; para poeira foi coletado um pool em área de risco, DR385 (385m) e em DR1700. Foi avaliada a atividade mutagênica através do ensaio Salmonella/microssoma, método de microssuspensão, em linhagens que detectam erro no quadro de leitura (TA98 e TA97a), substituição de pares de bases do DNA (TA100), em presença e ausência de metabolização hepática de ratos (S9mix) e YGs 1041, 1042 e 1024 para a definição de nitroderivados. Foram preparados extratos ácidos, visando definir os efeitos de compostos inorgânicos como metais pesados e de extratos orgânicos para avaliar principalmente hidrocarbonetos policíclicos aromáticos (HPAs) e nitroderivados. Os extratos foram testados também quanto à concentração dos 16 HPAs considerados poluentes prioritários pela USEPA, quanto a metais pesados de interesse e quanto ao contaminante pentaclorofenol (PCP) como marcador específico da atividade do sítio industrial. Através das análises de metais pesados foi detectado um gradiente de respostas, onde tanto a concentração de metais totais quanto a fração biodisponível de Cr e Cu apresentaram valores mais altos para SI em relação aos solos de entorno; para As, apenas na avaliação da concentração total do elemento, SI foi superior. Em relação aos efeitos mutagênicos decorrentes da mistura de compostos inorgânicos as respostas não permitiram uma gradação entre as diferentes distâncias. O solo fonte SI mostrou mutagênese nas diferentes linhagens, em especial em TA97a na ausência de S9mix. As amostras de entorno apresentaram potência mutagênica em pelo menos duas cepas, mas apenas uma consonante com SI. O solo SR500, mostrou mutagênese diferenciada e resposta expressiva na linhagem TA98 com S9 mix. No entanto, o local SR1700 mostrou ausência de influência a partir da área contaminada caracterizando uma área de referência. Nos extratos orgânicos, as respostas de mutagênese mostram um padrão de contaminação nas áreas de influência semelhante ao apresentado por SI, parecendo indicar a mobilidade de compostos orgânicos a partir da fonte para as áreas de entorno. Houve um predomínio de compostos de ação indireta, sendo os valores de SI entre 107 a 1455 rev/g de solo. Nos locais de entorno, observou-se padrão similar em SR150; já em SR500 valores elevados de mutagênese de ação direta foram evidenciados em TA97a; SR1700, embora apresentando mutagênese do tipo erro no quadro de leitura em presença de S9 mix, mostrou um decréscimo de efeitos. Os testes com as linhagens YG indicaram que compostos nitrados têm ação significativa na mutagênese direta encontrada, com exceção de SR500. Foi detectada ainda a presença de hidroxiamino-compostos em todas as amostras de solos através da linhagem YG1024. Na investigação da poeira residencial de entorno foram observadas respostas mutagênicas nas diferentes cepas testadas em DR385, mono e dinitroarenos do tipo substituição de pares de bases (YG1042) e hidroxiamino-compostos (YG1024); em DR1700 não foi observada resposta positiva. Concentrações de HPAs potencialmente carcinogênicos se estendem no solo da área interna até o do local de referencia, bem como na poeira domiciliar da área de risco indicando gradiente de concentrações e efeitos. A concentração de PCP no pool de poeira (DR385) foi (0,491 mg/Kg), similiar a observada em SI, (0,431 mg/Kg), definindo uma rota efetiva de dispersão a partir da área industrial para regiões do entorno. Ficou evidenciada a possibilidade de dispersão de mutágenos da área contaminada para regiões de entorno, sendo possível detectar gradientes de distância, favorecendo estimativas de risco. O estudo mostrou que é fundamental avaliar a extensão da contaminação a partir de fontes de solo impactado, visando delimitar qualquer medida de remediação dos ambientes atingidos e evitar danos potenciais ao equilíbrio ecológico e à saúde humana. / Soil is one of the compartments most affected by the accumulation of anthropic pollutants, since it acts as a deposit for pollutants and as a source for other interface environments. This study investigated a site contaminated with residues of the wood preservative industry as a source of mutagenic compounds; it defined the routes and area covered by dispersion of these contaminants through the remobilization of particles and atmospheric deposition, associating environmental and human health risks. Sampling sites were selected for soils and dusts at gradually increasing distances from SI (pool of soil from the industrial area), SR150 (150m), SR500 and SR1700; a pool was of residential dust was collected in the area of risk, DR385 (385m) and at DR1700. Mutagenic activity was evaluated by the Salmonella/microsome assay, microsuspension method, in strains that detect frameshift error (TA98 and TA97a), DNA base pair substitution (TA100), in the presence and absence of hepatic metabolization of rats (S9 mix) and YGs 1041, 1042 and 1024 to sough nitroderivates. Acid extracts were prepared to define the effects of inorganic compounds such as heavy metals, and organic extracts in evaluating mainly polycyclic aromatic hydrocarbons (PAHs) and nitroderivates. The extracts were tested also for the concentration of the 16 PAHs considered priority pollutants by USEPA, for heavy metals of interest and for contaminant pentachlorophenol (PCP) as a specific marker of the industrial site activity. A gradient of responses was detected through analyses of heavy metals , where both the concentration of heavy metals and the bioavailable fraction of Cr and Cu presented higher values for SI compared to the surrounding soils; for As, SI was superior only to evaluate the total concentration of the element. As to the mutagenic effects of the mixture of inorganic compounds, the responses did not allow a grading between the different distances. The source soil SI presented mutagenesis in the different strains, especially in TA97a in the absence of S9mix. The samples from the surrounding area presented mutagenic potency in at least two strains, but only one in accordance with SI. Soil SR500 showed differentiated mutagenesis and an expressive response in the TA98 strain with S9 mix. However, site SR1700 showed no influence from the contaminated area, characterizing an area of reference. In the organic extracts, the mutagenesis responses showed a pattern of contamination of the areas of influence similar to that presented by SI, and appear to indicate the mobility of organic compounds from the source to the surrounding areas. Indirect action compounds predominated, and the values of SI were from 107 to 1455 rev/g soil. At the surrounding sites, a similar pattern was observed in SR150; on the other hand, in SR500 high values of direct action mutagenesis were evidenced in TA97a; SR1700, although presenting mutagenesis in frameshift strains in the presence of S9 mix, showed diminished effects. Tests with YGs strains indicate that the nitrated compounds exert a significant action on the direct mutagenesis found, except for SR500. Further, hydroxyamine-compounds were detected in all soils samples through strain YG1024. Investigating residential dust from the surrounding area, mutagenic responses were observed in the different strains tested in DR385, mono and dinitroarenes of the pair substitution mutation type (YG1042) and hydroxyamine-compounds (YG1024); no positive response was observed in DR1700. Potentially carcinogenic PAHs concentrations are on the soil from the internal area until the site of reference, and also in the residential dust of the risk area, indicating a gradient of concentrations and effects. PCP concentration in the dust pool (DR385) was (0.491 mg/Kg), similar to that observed in SI, (0.431 mg/Kg), defining an effective dispersion route from the industrial area to the surrounding regions. The possibility of mutagen dispersion from the contaminated area to the surrounding regions was shown, and gradients of distance favoring risk estimates were detected. The study showed that it is essential to evaluate the extent of contamination from the sources of soil that have been impacted, with a view to delimiting any remedial measure for the environments affected and avoiding potential damage to the ecological balance and human health.

O solo é um dos compartimentos mais atingidos pelo acúmulo de poluentes de origem antrópica, pois atua como depósito de poluentes e como fonte para outros ambientes de interface. Este estudo investigou sítio contaminado com resíduos da indústria de preservativos de madeira como fonte de compostos mutagênicos; definiu rotas e abrangência na dispersão desses contaminantes pela remobilização de partículas e deposição atmosférica, associando riscos ambientais e à saúde. Foram selecionados locais de amostragem para solos e poeira domiciliar a distâncias gradativas a partir de SI (pool de solo da área industrial), SR150 (150m), SR500 e SR1700; para poeira foi coletado um pool em área de risco, DR385 (385m) e em DR1700. Foi avaliada a atividade mutagênica através do ensaio Salmonella/microssoma, método de microssuspensão, em linhagens que detectam erro no quadro de leitura (TA98 e TA97a), substituição de pares de bases do DNA (TA100), em presença e ausência de metabolização hepática de ratos (S9mix) e YGs 1041, 1042 e 1024 para a definição de nitroderivados. Foram preparados extratos ácidos, visando definir os efeitos de compostos inorgânicos como metais pesados e de extratos orgânicos para avaliar principalmente hidrocarbonetos policíclicos aromáticos (HPAs) e nitroderivados. Os extratos foram testados também quanto à concentração dos 16 HPAs considerados poluentes prioritários pela USEPA, quanto a metais pesados de interesse e quanto ao contaminante pentaclorofenol (PCP) como marcador específico da atividade do sítio industrial. Através das análises de metais pesados foi detectado um gradiente de respostas, onde tanto a concentração de metais totais quanto a fração biodisponível de Cr e Cu apresentaram valores mais altos para SI em relação aos solos de entorno; para As, apenas na avaliação da concentração total do elemento, SI foi superior. Em relação aos efeitos mutagênicos decorrentes da mistura de compostos inorgânicos as respostas não permitiram uma gradação entre as diferentes distâncias. O solo fonte SI mostrou mutagênese nas diferentes linhagens, em especial em TA97a na ausência de S9mix. As amostras de entorno apresentaram potência mutagênica em pelo menos duas cepas, mas apenas uma consonante com SI. O solo SR500, mostrou mutagênese diferenciada e resposta expressiva na linhagem TA98 com S9 mix. No entanto, o local SR1700 mostrou ausência de influência a partir da área contaminada caracterizando uma área de referência. Nos extratos orgânicos, as respostas de mutagênese mostram um padrão de contaminação nas áreas de influência semelhante ao apresentado por SI, parecendo indicar a mobilidade de compostos orgânicos a partir da fonte para as áreas de entorno. Houve um predomínio de compostos de ação indireta, sendo os valores de SI entre 107 a 1455 rev/g de solo. Nos locais de entorno, observou-se padrão similar em SR150; já em SR500 valores elevados de mutagênese de ação direta foram evidenciados em TA97a; SR1700, embora apresentando mutagênese do tipo erro no quadro de leitura em presença de S9 mix, mostrou um decréscimo de efeitos. Os testes com as linhagens YG indicaram que compostos nitrados têm ação significativa na mutagênese direta encontrada, com exceção de SR500. Foi detectada ainda a presença de hidroxiamino-compostos em todas as amostras de solos através da linhagem YG1024. Na investigação da poeira residencial de entorno foram observadas respostas mutagênicas nas diferentes cepas testadas em DR385, mono e dinitroarenos do tipo substituição de pares de bases (YG1042) e hidroxiamino-compostos (YG1024); em DR1700 não foi observada resposta positiva. Concentrações de HPAs potencialmente carcinogênicos se estendem no solo da área interna até o do local de referencia, bem como na poeira domiciliar da área de risco indicando gradiente de concentrações e efeitos. A concentração de PCP no pool de poeira (DR385) foi (0,491 mg/Kg), similiar a observada em SI, (0,431 mg/Kg), definindo uma rota efetiva de dispersão a partir da área industrial para regiões do entorno. Ficou evidenciada a possibilidade de dispersão de mutágenos da área contaminada para regiões de entorno, sendo possível detectar gradientes de distância, favorecendo estimativas de risco. O estudo mostrou que é fundamental avaliar a extensão da contaminação a partir de fontes de solo impactado, visando delimitar qualquer medida de remediação dos ambientes atingidos e evitar danos potenciais ao equilíbrio ecológico e à saúde humana. / Soil is one of the compartments most affected by the accumulation of anthropic pollutants, since it acts as a deposit for pollutants and as a source for other interface environments. This study investigated a site contaminated with residues of the wood preservative industry as a source of mutagenic compounds; it defined the routes and area covered by dispersion of these contaminants through the remobilization of particles and atmospheric deposition, associating environmental and human health risks. Sampling sites were selected for soils and dusts at gradually increasing distances from SI (pool of soil from the industrial area), SR150 (150m), SR500 and SR1700; a pool was of residential dust was collected in the area of risk, DR385 (385m) and at DR1700. Mutagenic activity was evaluated by the Salmonella/microsome assay, microsuspension method, in strains that detect frameshift error (TA98 and TA97a), DNA base pair substitution (TA100), in the presence and absence of hepatic metabolization of rats (S9 mix) and YGs 1041, 1042 and 1024 to sough nitroderivates. Acid extracts were prepared to define the effects of inorganic compounds such as heavy metals, and organic extracts in evaluating mainly polycyclic aromatic hydrocarbons (PAHs) and nitroderivates. The extracts were tested also for the concentration of the 16 PAHs considered priority pollutants by USEPA, for heavy metals of interest and for contaminant pentachlorophenol (PCP) as a specific marker of the industrial site activity. A gradient of responses was detected through analyses of heavy metals , where both the concentration of heavy metals and the bioavailable fraction of Cr and Cu presented higher values for SI compared to the surrounding soils; for As, SI was superior only to evaluate the total concentration of the element. As to the mutagenic effects of the mixture of inorganic compounds, the responses did not allow a grading between the different distances. The source soil SI presented mutagenesis in the different strains, especially in TA97a in the absence of S9mix. The samples from the surrounding area presented mutagenic potency in at least two strains, but only one in accordance with SI. Soil SR500 showed differentiated mutagenesis and an expressive response in the TA98 strain with S9 mix. However, site SR1700 showed no influence from the contaminated area, characterizing an area of reference. In the organic extracts, the mutagenesis responses showed a pattern of contamination of the areas of influence similar to that presented by SI, and appear to indicate the mobility of organic compounds from the source to the surrounding areas. Indirect action compounds predominated, and the values of SI were from 107 to 1455 rev/g soil. At the surrounding sites, a similar pattern was observed in SR150; on the other hand, in SR500 high values of direct action mutagenesis were evidenced in TA97a; SR1700, although presenting mutagenesis in frameshift strains in the presence of S9 mix, showed diminished effects. Tests with YGs strains indicate that the nitrated compounds exert a significant action on the direct mutagenesis found, except for SR500. Further, hydroxyamine-compounds were detected in all soils samples through strain YG1024. Investigating residential dust from the surrounding area, mutagenic responses were observed in the different strains tested in DR385, mono and dinitroarenes of the pair substitution mutation type (YG1042) and hydroxyamine-compounds (YG1024); no positive response was observed in DR1700. Potentially carcinogenic PAHs concentrations are on the soil from the internal area until the site of reference, and also in the residential dust of the risk area, indicating a gradient of concentrations and effects. PCP concentration in the dust pool (DR385) was (0.491 mg/Kg), similar to that observed in SI, (0.431 mg/Kg), defining an effective dispersion route from the industrial area to the surrounding regions. The possibility of mutagen dispersion from the contaminated area to the surrounding regions was shown, and gradients of distance favoring risk estimates were detected. The study showed that it is essential to evaluate the extent of contamination from the sources of soil that have been impacted, with a view to delimiting any remedial measure for the environments affected and avoiding potential damage to the ecological balance and human health.

O solo é um dos compartimentos mais atingidos pelo acúmulo de poluentes de origem antrópica, pois atua como depósito de poluentes e como fonte para outros ambientes de interface. Este estudo investigou sítio contaminado com resíduos da indústria de preservativos de madeira como fonte de compostos mutagênicos; definiu rotas e abrangência na dispersão desses contaminantes pela remobilização de partículas e deposição atmosférica, associando riscos ambientais e à saúde. Foram selecionados locais de amostragem para solos e poeira domiciliar a distâncias gradativas a partir de SI (pool de solo da área industrial), SR150 (150m), SR500 e SR1700; para poeira foi coletado um pool em área de risco, DR385 (385m) e em DR1700. Foi avaliada a atividade mutagênica através do ensaio Salmonella/microssoma, método de microssuspensão, em linhagens que detectam erro no quadro de leitura (TA98 e TA97a), substituição de pares de bases do DNA (TA100), em presença e ausência de metabolização hepática de ratos (S9mix) e YGs 1041, 1042 e 1024 para a definição de nitroderivados. Foram preparados extratos ácidos, visando definir os efeitos de compostos inorgânicos como metais pesados e de extratos orgânicos para avaliar principalmente hidrocarbonetos policíclicos aromáticos (HPAs) e nitroderivados. Os extratos foram testados também quanto à concentração dos 16 HPAs considerados poluentes prioritários pela USEPA, quanto a metais pesados de interesse e quanto ao contaminante pentaclorofenol (PCP) como marcador específico da atividade do sítio industrial. Através das análises de metais pesados foi detectado um gradiente de respostas, onde tanto a concentração de metais totais quanto a fração biodisponível de Cr e Cu apresentaram valores mais altos para SI em relação aos solos de entorno; para As, apenas na avaliação da concentração total do elemento, SI foi superior. Em relação aos efeitos mutagênicos decorrentes da mistura de compostos inorgânicos as respostas não permitiram uma gradação entre as diferentes distâncias. O solo fonte SI mostrou mutagênese nas diferentes linhagens, em especial em TA97a na ausência de S9mix. As amostras de entorno apresentaram potência mutagênica em pelo menos duas cepas, mas apenas uma consonante com SI. O solo SR500, mostrou mutagênese diferenciada e resposta expressiva na linhagem TA98 com S9 mix. No entanto, o local SR1700 mostrou ausência de influência a partir da área contaminada caracterizando uma área de referência. Nos extratos orgânicos, as respostas de mutagênese mostram um padrão de contaminação nas áreas de influência semelhante ao apresentado por SI, parecendo indicar a mobilidade de compostos orgânicos a partir da fonte para as áreas de entorno. Houve um predomínio de compostos de ação indireta, sendo os valores de SI entre 107 a 1455 rev/g de solo. Nos locais de entorno, observou-se padrão similar em SR150; já em SR500 valores elevados de mutagênese de ação direta foram evidenciados em TA97a; SR1700, embora apresentando mutagênese do tipo erro no quadro de leitura em presença de S9 mix, mostrou um decréscimo de efeitos. Os testes com as linhagens YG indicaram que compostos nitrados têm ação significativa na mutagênese direta encontrada, com exceção de SR500. Foi detectada ainda a presença de hidroxiamino-compostos em todas as amostras de solos através da linhagem YG1024. Na investigação da poeira residencial de entorno foram observadas respostas mutagênicas nas diferentes cepas testadas em DR385, mono e dinitroarenos do tipo substituição de pares de bases (YG1042) e hidroxiamino-compostos (YG1024); em DR1700 não foi observada resposta positiva. Concentrações de HPAs potencialmente carcinogênicos se estendem no solo da área interna até o do local de referencia, bem como na poeira domiciliar da área de risco indicando gradiente de concentrações e efeitos. A concentração de PCP no pool de poeira (DR385) foi (0,491 mg/Kg), similiar a observada em SI, (0,431 mg/Kg), definindo uma rota efetiva de dispersão a partir da área industrial para regiões do entorno. Ficou evidenciada a possibilidade de dispersão de mutágenos da área contaminada para regiões de entorno, sendo possível detectar gradientes de distância, favorecendo estimativas de risco. O estudo mostrou que é fundamental avaliar a extensão da contaminação a partir de fontes de solo impactado, visando delimitar qualquer medida de remediação dos ambientes atingidos e evitar danos potenciais ao equilíbrio ecológico e à saúde humana. / Soil is one of the compartments most affected by the accumulation of anthropic pollutants, since it acts as a deposit for pollutants and as a source for other interface environments. This study investigated a site contaminated with residues of the wood preservative industry as a source of mutagenic compounds; it defined the routes and area covered by dispersion of these contaminants through the remobilization of particles and atmospheric deposition, associating environmental and human health risks. Sampling sites were selected for soils and dusts at gradually increasing distances from SI (pool of soil from the industrial area), SR150 (150m), SR500 and SR1700; a pool was of residential dust was collected in the area of risk, DR385 (385m) and at DR1700. Mutagenic activity was evaluated by the Salmonella/microsome assay, microsuspension method, in strains that detect frameshift error (TA98 and TA97a), DNA base pair substitution (TA100), in the presence and absence of hepatic metabolization of rats (S9 mix) and YGs 1041, 1042 and 1024 to sough nitroderivates. Acid extracts were prepared to define the effects of inorganic compounds such as heavy metals, and organic extracts in evaluating mainly polycyclic aromatic hydrocarbons (PAHs) and nitroderivates. The extracts were tested also for the concentration of the 16 PAHs considered priority pollutants by USEPA, for heavy metals of interest and for contaminant pentachlorophenol (PCP) as a specific marker of the industrial site activity. A gradient of responses was detected through analyses of heavy metals , where both the concentration of heavy metals and the bioavailable fraction of Cr and Cu presented higher values for SI compared to the surrounding soils; for As, SI was superior only to evaluate the total concentration of the element. As to the mutagenic effects of the mixture of inorganic compounds, the responses did not allow a grading between the different distances. The source soil SI presented mutagenesis in the different strains, especially in TA97a in the absence of S9mix. The samples from the surrounding area presented mutagenic potency in at least two strains, but only one in accordance with SI. Soil SR500 showed differentiated mutagenesis and an expressive response in the TA98 strain with S9 mix. However, site SR1700 showed no influence from the contaminated area, characterizing an area of reference. In the organic extracts, the mutagenesis responses showed a pattern of contamination of the areas of influence similar to that presented by SI, and appear to indicate the mobility of organic compounds from the source to the surrounding areas. Indirect action compounds predominated, and the values of SI were from 107 to 1455 rev/g soil. At the surrounding sites, a similar pattern was observed in SR150; on the other hand, in SR500 high values of direct action mutagenesis were evidenced in TA97a; SR1700, although presenting mutagenesis in frameshift strains in the presence of S9 mix, showed diminished effects. Tests with YGs strains indicate that the nitrated compounds exert a significant action on the direct mutagenesis found, except for SR500. Further, hydroxyamine-compounds were detected in all soils samples through strain YG1024. Investigating residential dust from the surrounding area, mutagenic responses were observed in the different strains tested in DR385, mono and dinitroarenes of the pair substitution mutation type (YG1042) and hydroxyamine-compounds (YG1024); no positive response was observed in DR1700. Potentially carcinogenic PAHs concentrations are on the soil from the internal area until the site of reference, and also in the residential dust of the risk area, indicating a gradient of concentrations and effects. PCP concentration in the dust pool (DR385) was (0.491 mg/Kg), similar to that observed in SI, (0.431 mg/Kg), defining an effective dispersion route from the industrial area to the surrounding regions. The possibility of mutagen dispersion from the contaminated area to the surrounding regions was shown, and gradients of distance favoring risk estimates were detected. The study showed that it is essential to evaluate the extent of contamination from the sources of soil that have been impacted, with a view to delimiting any remedial measure for the environments affected and avoiding potential damage to the ecological balance and human health.

Orientadores: Antonio Jose Gomez Cobo, Inmaculada Rodriguez Ramos / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Quimica / Made available in DSpace on 2018-08-09T18:45:47Z (GMT). No. of bitstreams: 1 Zonetti_PrisciladaCosta_D.pdf: 1969256 bytes, checksum: 26749d2eefdd647b4142060303484744 (MD5) Previous issue date: 2007 / Resumo: Os compostos organoclorados são substâncias químicas que têm como característica uma elevada toxicidade e persistência, tanto no meio ambiente quanto nos organismos vivos. Tal é o caso do pentaclorofenol, usado como herbicida e pesticida na proteção de lavouras e como biocida no tratamento de madeiras e couros. Um método de tratamento bastante promissor é a hidrodescloração catalítica desses compostos empregando catalisadores à base de paládio. Dentro desse contexto, o presente trabalho tem como objetivo principal estudar as condições de preparação de catalisadores Pd/TiO2 e o comportamento dos sistemas bimetálicos Pd-Ni/TiO2, Pd-Ru/TiO2 e Pd-Ti/TiO2. Dentre as variáveis de preparação dos catalisadores Pd/TiO2, foram estudadas as influências do tratamento de ativação, bem como do teor de Pd no sólido sobre o desempenho catalítico. Para tanto, os catalisadores de Pd/TiO2 foram preparados pelo método de impregnação a seco e os catalisadores bimetálicos foram preparados pelo método de co-impregnação. Os sólidos obtidos foram caracterizados por meio das técnicas de adsorção de N2 (BET), análise espectrométrica de raios X (EDX), espectroscopia fotoeletrônica de raios X (XPS), redução à temperatura programada (RTP), espectroscopia no infravermelho da adsorção do CO (FTIR) e microcalorimetria da adsorção do CO. O desempenho dos catalisadores foi avaliado na reação de hidrodescloração do pentaclorofenol, empregandose um reator Parr do tipo "slurry". A reação foi conduzida à pressão de 5 bar e temperatura de 383 K. Os resultados dos testes catalíticos mostram que o catalisador 3%Pd/TiO2 não calcinado e não reduzido permite obter maiores valores para o rendimento e a seletividade de fenol, com uma atividade catalítica relativamente elevada. Os catalisadores bimetálicos com Ni e Ti são seletivos à formação de fenol, enquanto que o catalisador contendo Ru se mostra muito seletivo para a obtenção do cicloexanol, através da hidrogenação do anel benzênico / Abstract: Organic-chlorine compounds are chemicals which main characteristic is high toxicity and persistence, both in live organisms and in the environment. Such is the case of pentachlorophenol, used as herbicide and pesticide for harvest protection and as biocide for treating woods and leathers. A very promising treatment method is the catalytic hydrodechlorination of these products using palladium based catalysts. In this context, the following work is aimed at studying the Pd/TiO2 preparation conditions and the behavior of bimetallic Pd-Ni/TiO2, Pd-Ru/TiO2 and Pd-Ti/TiO2 systems. Within the preparation variables of Pd/TiO2 catalysts, the effects of the activation treatment as well as the influence of Pd content in the solid on the catalysts performance were studied. Therefore, Pd/TiO2 catalysts were prepared by means of the dry impregnation method, and the bimetallic catalysts were prepared by co-impregnation method. The so obtained solids were characterized by the N2 adsorption technique (BET), Xray spectrometry analysis (EDX), X-ray photoelectron spectroscopy (XPS), temperatureprogrammed reduction (TPR), Fourier transformed-infrared spectroscopy of the chemisorbed CO (FTIR) and CO adsorption microcalorimetry. Catalysts performance was evaluated in the entachlorophenol hydrodechlorination reaction using a slurry Parr reactor. The reaction was conducted at 5 bar pressure and at 383 K temperature. Catalytic test results show that the non calcinated and non reduced 3% Pd/TiO2 catalyst allows to attain higher values for yield and for phenol selectivity, with a relatively high catalytic activity. Bimetallic catalysts with Ni and Ti are selective in the phenol formation, while the catalyst with Ru is likely more selective to attain the cyclohexanol by means of the benzene ring hydrogenation / Doutorado / Sistemas de Processos Quimicos e Informatica / Doutor em Engenharia Química

Orientador : Antonio Jose Gomez Cobo / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Quimica / Made available in DSpace on 2018-08-04T00:25:53Z (GMT). No. of bitstreams: 1 Silva_JoseWilsonda_D.pdf: 3310228 bytes, checksum: fc913616b3b5d361419a1d4d39107d50 (MD5) Previous issue date: 2004 / Resumo: Desde a revolução industrial até os dias de hoje, a poluição do meio ambiente passou de um simples reflexo do progresso do homem, para ser um sério problema global. Atualmente, são várias as legislações que tentam restringir essa poluição, sendo esse fato o responsável pelo urgimento de várias tecnologias para o tratamento prévio dos resíduos tóxicos a serem lançados no meio ambiente. Uma dessas tecnologias é a hidrodescloração catalítica. Dentro desse contexto, o objetivo do presente estudo é a avaliação das condições experimentais para a hidrodescloração do pentaclorofenol com catalisadores de paládio. Essa avaliação foi realizada por meio de testes catalíticos com os catalisadores 5%Pd/C e 5%Pd/TiO2. Esses testes foram feitos de uma forma planejada, sendo que para tal empregou-se o planejamento fatorial fracionário 2 6/2. No que se refere aos catalisadores empregados, o sólido 5%Pd/C é um catalisador comercial, enquanto o catalisador 5%Pd/TiO2 foi preparado via impregnação a seco, partindo-se de uma solução aquosa de PdCl2. Depois de impregnado e seco, esse sólido foi dividido em duas partes: uma delas foi calcinada a 300 °C e a outra reduzida à mesma temperatura. Os resultados dos testes catalíticos revelaram que os catalisadores 5%Pd/C e 5%Pd/TiO2 reduzidos não foram ativos para a hidrodescloração do pentaclorofenol em fase líquida. Por outro lado, esses mesmos catalisadores, na sua forma oxidada, não foram desativados durante o tempo de reação em que foram avaliados. Além disso, esses testes catalíticos também revelaram que o catalisador 5%Pd/C não ativado é seletivo a cicloexanol, enquanto que o catalisador 5%Pd/TiO2 calcinado é seletivo a fenol / Abstract: Concerns with environmental pollution have changed most intensively since Industrial Revolution. At the beginning, environmental pollution was faced as a simple effect of the progress of humanity, but lately, it has been considered a terrible global problem. Nowadays, severe legislation for environmental pollution control has given rise to a great variety of technologies for pollution treatment. One of the solutions being proposed is catalytic hydrodechlorination. Taking this in consideration, the aim of this work is to evaluate the reaction conditions for pentachlorophenol hydrodechlorination over palladium based catalysts. This evaluation was derived from catalytic tests using 5%Pd/C and 5%Pd/TiO2 catalysts. These tests were done according to a fractional fatorial design 26-2. The 5%Pd/C solid is a comercial catalyst while the 5%Pd/TiO2 catalyst was prepared by incipient wetness impregnation technique from an aqueous solution of PdCl2. After drying, this solid was divided into two parts: one fo them was calcined 300 °C and the another was only reduced 300 °C. Results of catalyst tests indicated that 5%Pd/C and 5%Pd/TiO2 both reduced catalysts do not show any catalytic activity for liquid phase entachlorophenol hydrodechlorination. On the other hand, under an oxide form, these catalysts were not deactivated during the reaction time under investigation. Moreover, these catalyst performance also showed that the 5%Pd/C not reduced catalyst is selective to cyclohexanol, while the 5%Pd/TiO2 calcined catalyst is selective to phenol / Doutorado / Sistema de Processos Quimicos e Informatica / Doutor em Engenharia Química

Natural microbial communities are the ultimate drivers of change in any ecosystem. Through chemical contamination of natural environments, these communities are exposed to many different types of chemical stressors; however, research on whole genome responses to this contaminant stress is limited. This research examined the stress response of a common soil bacterium, <i>Pseudomonas aeruginosa</i>, to a common environmental pollutant, pentachlorophenol (PCP). In the first part of the research, it was revealed that nutrient-limited <i>P. aeruginosa</i> is able to respond to PCP with minimal physiological damage due to the upregulation of multidrug efflux pumps. Further study of this PCP-mediated induction of efflux pumps revealed a simultaneous increase in antibiotic resistance. It was discovered that the resistance nodulation-cell division (RND) efflux pump, MexAB-OprM, in particular is responsible for the PCP-induced increase in antibiotic resistance. Both whole cell physiological indicators and whole genome analysis were used to examine the stress response of <i>P. aeruginosa</i> to PCP. Cells were grown in a chemostat at a low growth rate to simulate nutrient-limiting growth in the natural environment. Whole cell acetate uptake rates (WAUR) and viable cell counts as colony forming units (CFU) were determined as cells were exposed to increasing concentration of PCP. At the same time, changes in gene expression were examined by Affymetrix microarray technology. Results showed little change in whole-cell physiology, with no difference in WAUR and only a slight reduction in CFU. However, the microarrays revealed that over 100 genes either increased or decreased expression greater than two-fold due to the PCP exposure. In particular, multiple multidrug efflux genes were upregulated in response to the PCP. The results were validated by real time reverse transcription polymerase chain reaction (RT-PCR) for one of these genes. Further analysis of the effects of MexAB-OprM showed that this particular efflux pump is essential for the response of <i>P. aeruginosa</i> to the toxin PCP. Induction of multidrug efflux pumps is responsible for the development of antibiotic resistance in strains of <i>P. aeruginosa</i>. Therefore, it was investigated whether PCP might induce resistance to a variety of antibiotics. The research was further extended to examine the effect of a variety of organic contaminants on MexAB-OprM efflux and antibiotic resistance development. PCP, 2,4-dinitrophenol, benzoate and Roundup® all induced antibiotic resistance. However, although MexAB-OprM is required for optimal growth in the presence of all chemicals, this particular efflux pump is only involved in increased resistance with PCP. This was confirmed using RT-PCR as <i>mexB</i> expression was induced by PCP, but not by the other three chemicals. A long term generational study on the effects of PCP did not result in a stable antibiotic-resistant phenotype; however, RT-PCR showed that <i>mexB</i> induction is a direct result of PCP exposure and can be reversed by removal of PCP. Together, these results demonstrate the necessity to understand functional responses to contaminant stress. Discovery of direct induction of multidrug efflux pumps and the resulting increase in antibiotic resistance has significant implications for environmental microbiology and public health. This research suggests that organic contamination may result in antibiotic resistance and that antibiotic resistant strains may have a survival advantage in contaminated environments. / Ph. D.