A conservação da biodiversidade edáfica na Floresta Ombrófila Mista favorece os processos de decomposição e ciclagem de nutrientes, melhorando as propriedades físicoquímicas e biológicas do solo. O objetivo deste trabalho foi verificar o potencial dos atributos da macrofauna edáfica, Fungos Micorrízicos Arbusculares (FMA) e de variáveis ambientais explicativas (físico-químicas e microbiológicas) como indicadores de qualidade do solo em duas épocas contrastantes (inverno e verão) em Floresta de Araucária do Estado de São Paulo. Foram avaliadas florestas com Araucaria angustifolia nativa (NF) e reflorestada (RF) em três regiões distintas do Estado de São Paulo. Cada floresta teve três repetições verdadeiras, nas quais, foram coletadas, ao acaso, quinze amostras de solo para avaliação dos atributos físicos (densidade, macroporosidade, microporosidade, porosidade total e umidade), químicos [pH, teores de carbono orgânico (C-org), P, Ca, Mg, K, Al e H+Al] e biológicos [macrofauna, FMA, carbono da biomassa microbiana (CBM), respiração basal, quociente metabólico (qCO2) e atividade da desidrogenase (Desi)]. Nos mesmos pontos de amostragem, foram avaliadas a macrofauna edáfica, a densidade de FMA e a colonização micorrízica em raízes de araucária. Na serapilheira foi avaliada a massa seca (MSS) e foram determinados os teores de C, N e S. Os atributos físico-químicos e microbiológicos do solo e químicos da serapilheira foram submetidos à análise de variância (Two-way ANOVA). A abundância da macrofauna e de espécies de minhocas foram submetidas à Análise de Componentes Principais (ACP) e as variáveis microbiológicas, MSS, físico-químicas do solo e da serapilheira foram usadas como variáveis explicativas (VE). A abundância de espécies de FMA foi submetida à ACP e as variáveis físico-químicas do solo e da serapilheira foram usadas como VE. Adicionalmente, a abundância da macrofauna e as VE foram submetidas à Análise Canônica Discriminante (ACD) e a uma Análise de Correlação Canônica (ACC). A macrofauna apresentou maior riqueza de grupos no verão, sendo que a NF proporcionou a maior diversidade de grupos. A umidade, porosidade total, teor de S na serapilheira, pH, teor de K, CBM e respiração basal foram os atributos edáficos mais importantes para separar as áreas. O grupo Oligochaeta, principamente as espécies Ponthoscolex corethrurus e Amynthas spp., foram boas indicadoras de qualidade do solo e/ou de distúrbio na Floresta de Araucária. Encontraram-se 36 táxons de FMA, sendo os gêneros Glomus, Acaulospora e Scutellospora os mais abundantes. A espécie Ambispora appendicula apresentou a maior abundância relativa. No inverno, observou-se alta correlação da Desi e C-CO2 com pH e H+Al, enquanto que no verão, o C-org, umidade, Mg, pH e C da serapilheira se correlacionaram com a atividade da Desi e C-CO2. Os atributos H+Al, P, porosidade total, S da serapilheira e umidade do solo foram os mais importantes na separação das áreas, independente de épocas de coleta. / The conservation of edaphic biodiversity in Araucaria Forests favors the processes of decomposition and nutrient cycling, improving the physico-chemical and biological soil properties. The objective of this study was to investigate the potential of the macrofauna, mycorrhizal fungi (AMF) and environmental explanatory variables (physico, chemical and microbiological) as indicators of soil quality in two different contrasting seasons (winter and summer) in Araucaria Forests. We evaluated forests with native Araucaria angustifolia (NF) and reforested areas (RF) in three different regions in the State of São Paulo. Each forest had three true replicates, in which we collected at random, fifteen soil samples for evaluation of physical attributes (density, macroporosity, microporosity, total porosity and moisture), chemical attributes [pH, organic carbon (org-C), P, Ca, Mg, K, Al and H+Al] and biological attributes [macrofauna, AMF, microbial biomass carbon (MBC), basal respiration (CO2-C), metabolic quotient (qCO2) and dehydrogenase activity (Desi)]. At the same sampling points we evaluated the edaphic macrofauna, the density of AMF and root colonization in Araucaria. We evaluated the litter dry matter (MSS) and analyzed the contents of C, N and S. The physico-chemical and microbiological soil attributes and the chemical litter attributes were submitted to analysis of variance (Two-way ANOVA). The abundance of macrofauna and of earthworm species were submitted to a Principal Components Analysis (PCA) and the microbiological variables, litter dry weight, physico-chemical attributes of soil and litter were used as explanatory variables (EV). The abundance of AMF species was submitted to the PCA and the physico-chemical variables of soil and litter were used as EV. Additionally, the abundance of macrofauna and EV were submitted to a Canonical Discriminant Analysis (CDA) and to one Canonical Correlation Analysis (CCA). The macrofauna showed greater richness of groups in the summer, and the NF provided the highest diversity. Moisture, total porosity, S litter content, pH, K, MBC and basal respiration were the edaphic attributes most important to separate the areas. The group Oligochaeta, especially the species Ponthoscolex corethrurus and Amynthas spp., were good indicators of soil quality and/or disturbance in these Araucaria forests. We found 35 AMF taxons, and the genera Glomus, Acaulospora and Scutellospora were the most abundant. The species Ambispora appendicula showed the highest relative abundance. In winter, a high correlation of the microbiological attributes Desi and CO2-C with pH and H+Al was found, while in the summer, the org-C, moisture, Mg, pH and Carbon of the litter were more correlated with Desi activity and C-CO2. The attributes H+Al, P, total porosity, S of the litter and soil moisture were the most important attributes to discriminate the areas, regardless of the season.

A preocupação mundial com as possíveis mudanças climáticas, decorrentes do aumento da concentração de gases do efeito estufa (GEE), resultam na busca por fontes de energia renovável. Dentre estas fontes, a produção do etanol vem se destacando globalmente. O Brasil é o maior produtor mundial de etanol, baseado na cana-de-açúcar como matéria-prima. A produção deste biocombustível atua na redução das emissões de GEE, como substituinte de derivados de petróleo e fornecimento de energia, através da queima do bagaço. No entanto, é necessário quantificar o off-set do etanol em relação ao petróleo em todas as fases de produção. Importante resíduo da indústria sucroenergética é a vinhaça, gerada em elevadas quantidades e usualmente aplicada no solo em fertirrigação. Muitos estudos relatam os benefícios que a vinhaça proporciona ao solo, porém, pouco se sabe sobre os possíveis impactos que causa no ambiente, sobretudo no que se refere às emissões de GEE. O objetivo desta pesquisa foi avaliar as emissões de GEE provenientes da vinhaça no canal de distribuição e após aplicação no solo e detectar as comunidades de archaeas presentes no sedimento do canal condutor, verificando a influência de fatores bióticos e abióticos nas emissões e comunidades microbianas. A pesquisa foi desenvolvida na Usina Iracema, localizada em Iracemápolis, SP. Amostragens de GEE foram realizadas em diferentes pontos do canal de distribuição e em áreas de cana queimada e crua. Amostras de sedimento foram coletadas no canal para detecção das comunidades de archaeas por DGGE. As emissões de C-CO2 e C-CH4 provenientes da vinhaça no canal de distribuição foram influenciadas pela composição físico-química deste resíduo. Apesar de apresentar emissões significativas, o C-CO2 não foi computado no cálculo das emissões totais, pois é reassimilado pela fotossíntese. As emissões de N-N2O foram muito baixas, demonstrando que a vinhaça não fornece condições favoráveis à formação deste GEE. Ao converter os fluxos de N-N2O e C-CH4 em CO2 eq. observou-se que o C-CH4 contribuiu com aproximadamente 99% das emissões totais oriundas da vinhaça presente no canal. A fertirrigação do solo potencializou as emissões de CCO2 e N-N2O e não influenciou as emissões de C-CH4. As emissões de N-N2O provenientes da aplicação de nitrogênio na forma de vinhaça no solo resultaram em fatores de emissão de 0,68 e 0,44 % (kg N-N2O/kg N), respectivamente para cana queimada e crua. Após conversão dos fluxos de N-N2O e C-CH4 em CO2 eq. verificou-se que a aplicação da vinhaça no solo aumentou as emissões em 46,0 e 30,9 kg de CO2 eq. ha-1, respectivamente para cana queimada e crua. Considerando a aplicação de 200 m3 ha-1, observou-se que a vinhaça emitiu 493,9 e 489,1 kg de CO2 eq. ha-1, respectivamente para cana queimada e crua. A contabilização das emissões totais de GEE oriundas da vinhaça indicou que 90% das emissões são resultantes do canal de distribuição. A técnica de DGGE demonstrou mudanças nas estruturas das comunidades de archaea total e mcrA de acordo com os pontos de amostragem, exibiu forte relação com as condições encontradas nesses pontos e com a produção de C-CH4. / The global growing concern about an eventual climate change due to the increasing atmospheric greenhouse gases (GHG) concentrations spur on the search for renewable energy sources. Among the possibilities, ethanol is getting global prominence. Brazil is the greatest producer of ethanol derived from sugar-cane. The production of this biofuel helps to reduce GHG emission replacing the use of petroleum and producing energy by bagasse burning. However, it is necessary to quantify its effective off-set analyzing all steps of production. The main residue of the sugar- ethanol industry is the vinasse, which is generated in high quantity and is usually applied to soil as fertirrigation. Several studies mention the benefits of vinasse application to soil cultivated with sugarcane, but few is known about possible impacts on environment, especially in what concerns GHG emissions. The objective of this work was to evaluate GHG emissions from the vinasse during its flux in distribution channels and after its application to soil by aspersion. Besides, the structure of archaea community of the sludge deposited at the channels bottom was investigated, evaluating the influence of biotic and abiotic factors on GHG emission rates and microbial community. The study was developed at Usina Iracema, located in Iracemápolis (SP). GHG samples were taken at six different points along the vinasse distribution channel and after its application at sites where sugar cane is harvest preceded or not by straw burning. Samples from sedimented sludge were collected from the channel and archaeas communities were assessed by DGGE analyses. Emissions rates of CO2-C and CH4-C from vinasse were influenced by the physical-chemical composition of this residue. Although significant emission rates of CO2- C have been measured, they were not included in total emission calculations, since this gas is reabsorbed by photosynthesis. N2O-N emissions were very low, demonstrating that the vinasse does not promote favorable conditions to its building. The conversion of N2O-N and CH4-C fluxes into CO2 eq demonstrated that methane represents about 99% of total emission of vinasse while into the distribution channel. The soil fertirrigation with vinasse increased CO2-C and N2ON emissions, but did not influence C-CH4 emission rate. Nitrous oxide originated by N content in vinasse applied to soil represented 0.68 and 0.44 % (kg N2O-N.kg N-1), respectively, from areas were harvest was or was not preceeded by burning. N2O-N and CH4-C fluxes converted to CO2 eq showed that vinasse application increased emissions by 46.0 e 30.9 kg of CO2 eq. ha-1 respectively, at areas were harvest was or was not preceded by burning. Considering that vinasse is usually applied in rates of 200 m3 ha-1 the areas were harvest was or was not preceded by burning could emit a total of 493.9 and 489.1 kg CO2 eq. ha-1 respectively. In the final account, 90% of total GHG emission from vinasse was originated from the distribution channel. The DGGE technique demonstrated changes in total and functional community structure of archaeal according to sampling point. They were also strongly correlated to environmental conditions end C-CH4 production.

O algodão transgênico Bollgard® (algodão Bt) contém o gene cry1Ac da bactéria Bacillus thuringiensis, que confere a planta resistência a Lepidopteros. A expressão deste gene na planta pode acarretar efeitos ecológicos adversos à microbiota do solo e da rizosfera. Em casa-de-vegetação, a comunidade bacteriana associada ao algodão Bt foi comparada a do algodão convencional, em dois tipos de solos e quatro estádios fenológicos. Amostras de rizosfera foram avaliadas por técnicas dependentes e independentes de cultivo. As técnicas de contagem de bactérias e DGGE permitiram observar os efeitos do algodão Bt na densidade e diversidade de Pseudomonas e bactérias totais, durante os estádios iniciais de desenvolvimento da planta. A toxina Cry foi detectada na rizosfera de algodão Bt, em todo ciclo da cultura. Nas fases de formação do botão floral e abertura das maçãs, a atividade microbiana foi maior na rizosfera do algodão Bt. Esses resultados indicam o potencial do ambiente rizosférico em reestabelecer à estrutura da comunidade bacterina após um impacto temporal. / The transgenic cotton Bollgard® (Bt cotton) contains the cry1Ac gene from the Bacillus thuringiensis bacterium, which confers the plant resistance against some insects. The expression of this gene in the plant can cause adverse ecological effects on soil and rhizosphere microbiota. In a greenhouse experiment, the bacterial community associate to Bt cotton was compared to non-transgenic parental cultivar plants, in two types of soil at different plant development stages. Rhizosphere communities were evaluated by culture-dependent and independent approaches. Results reveal the effect of the Bt cotton in the density and diversity of Pseudomonas and total bacteria, during initial plant development stages. The Cry toxin was detected in the rhizosphere of Bt cotton, during all plant cycle. In the phases of flower formation and fruit opening, the microbial activity was greater in the rhizosphere of Bt cotton. These results show the potential of the rhizosphere to reestablish the original structure of the bacterial community after a temporary impact.

The microbial communities found in soils are inherently heterogeneous and often exhibit spatial variations on a small scale. Becker et al. (2006) investigate this phenomenon and present statistical analyses to support their findings. In this project, alternative statistical methods and models are considered and employed in a re-analysis of the data from Becker. First, parametric nested random effects models are considered as an alternative to the nonparametric semivariogram models and kriging methods employed by Becker to analyze patterns of spatial variation. Second, multiple logistic regression models are employed to investigate factors influencing microbial community structure as an alternative to the simple logistic models used by Becker. Additionally, the microbial community profile data of Becker were unobservable at several points in the spatial grid. The Becker analysis assumes that the data are missing completely at random and as such have relatively little impact on inference. In this re-analysis, this assumption is investigated and it is shown that the pattern of missingness is correlated with both metabolic potential and spatial coordinates and thus provides useful information that was previously ignored by Becker. Multiple imputation methods are employed to incorporate the information present in the missing data pattern and results are compared with those of Becker.

Devido ao seu elevado teor de nutrientes e potencial de neutralização da acidez do solo, a utilização de lodos de curtume em áreas agrícolas tem sido uma alternativa para disposição e reciclagem desses resíduos. Por outro lado, o aumento do pH e do teor de nitrogênio amoniacal no solo, quando da aplicação superficial do lodo de curtume, podem favorecer a perda de nitrogênio (N) por volatilização da amônia (NH3). Altos teores de nitrogênio inorgânico no solo podem gerar efeitos negativos, principalmente quando a amonificação e nitrificação não são sincronizadas com a absorção pelas plantas, possibilitando lixiviação e conseqüente contaminação das águas subsuperficiais. Um experimento de campo foi instalado em Rolândia (PR) com os objetivos de avaliar a perda de nitrogênio por volatilização da amônia, as alterações em alguns atributos microbiológicos do solo envolvidos no ciclo do nitrogênio e carbono, a lixiviação de nitrogênio mineral no solo, a produtividade da cultura de milho e o efeito residual, após a aplicação de doses crescentes de lodo de curtume no solo. O delineamento experimental foi de blocos completos casualizados, com quatro repetições. As doses de lodo foram calculadas em função do teor de N total contido no lodo de curtume. Os tratamentos foram: controle, 120, 480, 840 e 1200 kg ha-1 de N total adicionados via lodo de curtume, correspondendo, respectivamente, a 0, 3,4, 13,5, 23,6 e 33,7 Mg ha-1 de lodo de curtume (base seca). O lodo foi aplicado na superfície de um Nitossolo Vermelho distroférrico, muito argiloso, onde permaneceu por 89 dias, período em que foi determinada a perda de amônia por volatilização. Em seqüência, o lodo foi incorporado, com posterior semeadura da cultura de milho. Nesse momento foi instalado um tratamento adicional (Tratamento agronômico - 120 kg ha-1 N via uréia). Após a colheita do milho, a aveia preta foi semeada em sistema direto, sendo conduzida até o estádio de florescimento. Durante todo o período experimental foram realizadas coletas de solo nas camadas de 0-10, 10-20, 20-40, 40-60 cm e solução do solo nos tratamentos controle, 120 e 1200 kg ha-1 N total via lodo, a 1,2 m de profundidade. A volatilização da amônia foi mais intensa nos 30 primeiros dias após a aplicação, decaindo depois desse período. A fração volatilizada como NH3 correspondeu, em média, a 17,5% do N total contido no lodo de curtume. A densidade dos grupos funcionais de microrganismos, respiração do solo e atividade enzimática apresentaram aumentos transientes após aplicação de lodo de curtume. Dentre os atributos avaliados, as enzimas glutaminase, urease e asparaginase mostraram maior atividade em resposta à aplicação de lodo de curtume. A dose 120 kg ha-1 de N via lodo de curtume não apresentou risco de contaminação do lençol freático, enquanto que na maior dose (1200 kg ha-1 N total) o teor de nitrato na solução do solo coletada a 1,2 m foi até 12 vezes maior que no controle. A dose de 521 kg ha-1 de N total proporcionou ganhos de produtividade de grãos de 13% e 11% em relação ao controle e ao tratamento agronômico, respectivamente. Foi observado efeito residual sobre a massa de matéria seca da parte aérea da aveia preta 390 dias após a aplicação do lodo. Ou seja: Doses elevadas de lodo de curtume (equivalentes a altas doses de N), aplicadas ao solo, podem resultar em poluição do ar e das águas subsuperficiais. / Due to its high nutrient content and its neutralizing potential for soil acidity, the utilization of tannery sludge in agricultural areas can be an alternative for its disposal and recycling. On the other hand, the pH and ammonia nitrogen content increase in soil as a result of surface tannery sludge application and may induce the loss of nitrogen (N) by ammonia (NH3) volatilization. The high N content in soils can generate negative effects, mainly when the organic N ammonification and nitrification does not coincide with the plant N uptake, allowing the nitrate to leach through the soil profile to the groundwater. A field experiment was installed in Rolândia (Paraná State, Brazil). The aims of this experiment were to evaluate the N loss through ammonia volatilization, changes in soil microbial attributes that are linked to the carbon and N cycles, mineral N leaching through the soil profile, corn yield and the residual effect as affected by tannery sludge application on the soil surface. A randomized complete block design was used with four replicates. The tannery sludge treatments were: control, 120, 480, 840 and 1200 kg ha-1 total N applied as tannery sludge, equivalent to 0, 3.4, 13.5, 23.6 and 33.7 Mg ha-1 tannery sludge (dry weight). The tannery sludge was surface applied on a clayey Rhodic Kandiudult. The tannery sludge was left on the soil surface for 89 days, and during this period the N loss through NH3 volatilization was determined. Afterwards, tannery sludge was incorporated and the corn was sown. At this moment, an additional treatment (agricultural treatment - 120 kg ha-1 N as urea) was applied. After the corn harvest, black oat was sown and carried on until flowering. Soil samples were taken at the following depths: 0-10, 10-20, 20-40 and 40-60 cm for all treatments. Soil solution was extracted at 1.2 m soil depth only in the control, 120 and 1200 kg ha-1 total N treatments. NH3 volatilization was more intense during the beginning of the experiment (30 days). The volatilized fraction as NH3 corresponded in average to 17.5% of the total N of the tannery sludge. The functional microorganism density, soil respiration and enzyme activity showed transient increases after tannery sludge application. The glutaminase, urease and asparaginase enzymes showed a more pronounced increase in activity in response to tannery sludge application. The lowest dose of 120 kg ha-1 total N did not show a contamination risk to the groundwater, while in the highest dose (1200 kg ha-1 total N), the amount of nitrate detected at 1.2 m was 12 times higher than in the control. The dose of 521 kg ha-1 total N corresponded to the maximum corn yield and caused increases of 13% and 11%, when compared to the control and to the agricultural treatment, respectively. Even after 390 days, a residual effect of tannery sludge application was observed as an increase in shoot dry matter of black oat. In short: it was demonstrated that high doses of tannery sludge (equivalent to high doses of total N), applied to soil, result in prohibitively high values of air and ground water pollution.

by Ho, Kai-Man. / Thesis submitted in: December 2004. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 158-179). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstracts --- p.ii / Contents --- p.vii / List of figures --- p.xiv / List of tables --- p.xvii / Abbreviations --- p.xx / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Poly cyclic Aromatic Hydrocarbons (PAHs) / Chapter 1.1.1 --- Characteristics of PAHs --- p.1 / Chapter 1.1.2 --- Sources of PAHs --- p.3 / Chapter 1.1.3 --- Environmental fates of PAHs --- p.5 / Chapter 1.1.4 --- Effects of PAHs on living organisms --- p.7 / Chapter 1.1.5 --- Summary --- p.10 / Chapter 1.2 --- Target PAH: Benzo[a]pyrene (BaP) --- p.10 / Chapter 1.3 --- PAH contamination in Hong Kong --- p.14 / Chapter 1.4 --- Remediation for PAHs contaminated soils --- p.15 / Chapter 1.4.1 --- Chemical/ Physical methods --- p.15 / Chapter 1.4.2 --- Bioremediation --- p.16 / Chapter 1.5 --- Biodegradation of PAHs by bacteria and fungi --- p.18 / Chapter 1.5.1 --- Tolerance and degradation --- p.18 / Chapter 1.5.2 --- Biodegradation of PAHs by microorganisms --- p.20 / Chapter 1.5.2.1 --- Bacteria --- p.22 / Chapter 1.5.2.2 --- Fungi --- p.23 / Chapter 1.6 --- Environmental standards --- p.26 / Chapter 1.7 --- Strategies of soil sampling and microbial isolation --- p.26 / Chapter 1.7.1 --- Contaminated soil vs. uncontaminated soil --- p.26 / Chapter 1.7.2 --- Native species vs. foreign species --- p.29 / Chapter 1.7.3 --- Background of the sampling areas --- p.30 / Chapter 1.7.3.1 --- North Tsing Yi shipyard --- p.30 / Chapter 1.7.3.2 --- "Tsam Chuk Wan, Sai Kung" --- p.33 / Chapter 1.8 --- Objectives of this study --- p.33 / Chapter 2. --- Materials and Methods --- p.35 / Chapter 2.1 --- Soil Collection --- p.35 / Chapter 2.1.1 --- Abandoned shipyard soil and its sediment soil --- p.35 / Chapter 2.1.2 --- "Tsam Chuk Wan, Sai Kung" --- p.35 / Chapter 2.2 --- Characterization of soils --- p.35 / Chapter 2.2.1 --- Sample preparation --- p.36 / Chapter 2.2.2 --- Soil pH --- p.36 / Chapter 2.2.3 --- Electrical conductivity --- p.36 / Chapter 2.2.4 --- Salinity --- p.36 / Chapter 2.2.5 --- Total organic carbon contents --- p.38 / Chapter 2.2.6 --- Metal analys --- p.is / Chapter 2.2.7. --- Oil and grease content --- p.38 / Chapter 2.2.8 --- Soil texture --- p.39 / Chapter 2.2.9 --- Available ammoniacal nitrogen and oxidized nitrogen --- p.39 / Chapter 2.2.10 --- Available Phosporus --- p.40 / Chapter 2.2.11 --- Total Nitrogen and total Phosporus --- p.40 / Chapter 2.2.12 --- Moisture / Chapter 2.2.13 --- DTPA-extractable metals --- p.41 / Chapter 2.2.14 --- Extraction of PAHs and organic pollutants --- p.41 / Chapter 2.2.14.1 --- Extraction procedures --- p.41 / Chapter 2.2.14.2 --- GC-MSD conditions --- p.41 / Chapter 2.2.14.3 --- Extraction efficiency --- p.43 / Chapter 2.2.15 --- Soil colour --- p.43 / Chapter 2.3 --- Screening and selection of microorganismms --- p.43 / Chapter 2.3.1 --- Isolation of potential BaP-degrading microorganisms --- p.44 / Chapter 2.3.1.1 --- Isolation of bacteria --- p.44 / Chapter 2.3.1.2 --- Isolation of fungi --- p.44 / Chapter 2.3.2 --- Cultures preserving microorganisms --- p.46 / Chapter 2.3.3 --- Screening and selection of microbes --- p.46 / Chapter 2.3.3.1 --- Bacteria --- p.46 / Chapter 2.3.3.2 --- Fungi --- p.46 / Chapter 2.3.4 --- Survival test --- p.47 / Chapter 2.3.5 --- Removal efficiency (RE) towards BaP by the microorganisms --- p.47 / Chapter 2.3.5.1 --- Bacteria --- p.47 / Chapter 2.3.5.2 --- Fungi --- p.48 / Chapter 2.3.6 --- Removal efficiency (RE) --- p.48 / Chapter 2.3.7 --- Relationship of absorbance of bacterial culture and bacterial biomass --- p.49 / Chapter 2.4 --- Identification of selected microorganisms --- p.49 / Chapter 2.4.1 --- Identification of bacterium --- p.49 / Chapter 2.4.1.1 --- 16S rDNA sequencing --- p.49 / Chapter 2.4.1.1.1 --- Primers --- p.49 / Chapter 2.4.1.1.2 --- DNA extraction --- p.51 / Chapter 2.4.1.1.3 --- Specific PCR --- p.51 / Chapter 2.4.1.1.4 --- Gel electrophoresis --- p.51 / Chapter 2.4.1.1.5 --- Purification of PCR products --- p.52 / Chapter 2.4.1.1.6 --- DNA sequencing --- p.52 / Chapter 2.4.1.2 --- Midi Sherlock® Microbial Identification System (MIDI) --- p.53 / Chapter 2.4.1.3 --- Biolog MicroLog´ёØ system (Biolog) --- p.55 / Chapter 2.4.2 --- Identification of fungi --- p.56 / Chapter 2.4.2.1 --- ITS DNA sequencing --- p.56 / Chapter 2.4.2.2 --- Observation under electronic microscope --- p.58 / Chapter 2.5 --- Growth curve of the microorganism --- p.58 / Chapter 2.5.1 --- Bacterium --- p.58 / Chapter 2.5.2 --- Fungi --- p.58 / Chapter 2.6 --- Preparation of Benzo[a]pyrene (BaP) stock solution --- p.58 / Chapter 2.7 --- Comparison of isolated bacterium and fungi --- p.60 / Chapter 2.8 --- Optimization of BaP degradation by selected fungus --- p.60 / Chapter 2.8.1 --- Preparation of straw compost inoculated with selected fungus --- p.60 / Chapter 2.8.2 --- Effect of incubation time --- p.61 / Chapter 2.8.3 --- Effect of initial BaP concentration --- p.61 / Chapter 2.8.4 --- Effect of inoculum size / Chapter 2.8.5 --- Effect of temperature --- p.61 / Chapter 2.8.6 --- Effect of soil pH --- p.62 / Chapter 2.8.7 --- Study of BaP degradation pathway by the microorganisms using GC-MSD --- p.62 / Chapter 2.9 --- Chitin Assay --- p.62 / Chapter 2.10 --- Enzyme assay --- p.63 / Chapter 2.10.1 --- Laccase assay --- p.63 / Chapter 2.10.2 --- Manganese peroxidase assay --- p.63 / Chapter 2.10.3 --- Lignin peroxidase assay --- p.64 / Chapter 2.11 --- Toxicity of treated soil --- p.64 / Chapter 2.12 --- Statistical analysis --- p.65 / Chapter 3. --- Results --- p.66 / Chapter 3.1 --- Soil Collection --- p.66 / Chapter 3.1.1 --- North Tsing Yi shipyard --- p.66 / Chapter 3.1.2 --- "Tsam Chuk Wan, Sai Kung" --- p.66 / Chapter 3.2 --- Characterization of soil samples --- p.71 / Chapter 3.3 --- Extraction efficiency of Benzo[a]pyrene --- p.79 / Chapter 3.4 --- Screening and selection of microorganisms --- p.79 / Chapter 3.4.1 --- Isolation ofpotential BaP-degrading microorganisms --- p.79 / Chapter 3.4.2 --- Screening and selection of microbes --- p.87 / Chapter 3.4.2.1 --- Bacteria --- p.87 / Chapter 3.4.2.2 --- Fungi --- p.93 / Chapter 3.4.3 --- Growth curve of the microorganisms --- p.95 / Chapter 3.4.3.1 --- Bacterium --- p.95 / Chapter 3.4.3.2 --- Fungi --- p.99 / Chapter 3.5 --- Comparison of isolated bacterium and fungi --- p.99 / Chapter 3.6 --- Identification of selected microorganisms --- p.102 / Chapter 3.6.1 --- Identification of bacterium --- p.103 / Chapter 3.6.1.1 --- 16S rDNA sequencing --- p.103 / Chapter 3.6.1.2 --- Midi Sherlock® Microbial Identification System (MIDI) --- p.103 / Chapter 3.6.1.3 --- Biolog MicroLog´ёØ system (Biolog) --- p.107 / Chapter 3.6.2 --- Identification of fungi --- p.107 / Chapter 3.6.2.1 --- ITS DNA sequencing --- p.107 / Chapter 3.6.2.2 --- Observation under electronic microscope --- p.113 / Chapter 3.7 --- Optimization of BaP degradation by the selected fungus: Trichoderma asperellum --- p.107 / Chapter 3.7.1 --- Effect of incubation time --- p.107 / Chapter 3.7.2 --- Effect of inoculum size --- p.113 / Chapter 3.7.3 --- Effect of initial BaP concentration --- p.113 / Chapter 3.7.4 --- Effect of soil pH --- p.113 / Chapter 3.7.5 --- Effect of temperature --- p.117 / Chapter 3.8 --- Determination of breakdown products of BaP by BaP-degrading microorganisms --- p.117 / Chapter 3.9 --- Enzyme assay --- p.117 / Chapter 3.10 --- Evaluation of toxicity by using indigenous bacteria --- p.121 / Chapter 4. --- Discussion --- p.128 / Chapter 4.1 --- Physico-chemical properties of soil samples --- p.128 / Chapter 4.2 --- Determination of BaP and other organic compounds --- p.131 / Chapter 4.3 --- Identification of the microorganisms --- p.132 / Chapter 4.3.1 --- Bacteria --- p.132 / Chapter 4.3.2 --- Fungi --- p.134 / Chapter 4.4 --- Biodegradation by BaP-degrading microorganisms --- p.135 / Chapter 4.4.1 --- Isolation and screening of BaP-degrading microorganisms --- p.135 / Chapter 4.4.2 --- Biodegradation of BaP --- p.137 / Chapter 4.4.2.1 --- Bacteria --- p.137 / Chapter 4.4.2.2 --- Fungi --- p.138 / Chapter 4.4.3 --- BaP degradation pathway --- p.140 / Chapter 4.4.3.1 --- Bacteria --- p.140 / Chapter 4.4.3.2 --- Fungi --- p.140 / Chapter 4.5 --- Optimization of PAH degradation by T. asperellum --- p.143 / Chapter 4.5.1 --- Effect of incubation time --- p.143 / Chapter 4.5.2 --- Effect of initial BaP concentration --- p.144 / Chapter 4.5.3 --- Effect of inoculum size fungus --- p.144 / Chapter 4.5.4 --- Effect of soil pH --- p.145 / Chapter 4.5.5 --- Effect of temperature --- p.146 / Chapter 4.6 --- Comparison the selected bacterium and fungi --- p.146 / Chapter 4.7 --- Evaluation of toxicity by using in indigenous bacteria --- p.148 / Chapter 4.8 --- Post treatment by crude enzyme of Pleurotus pulmonarius --- p.149 / Chapter 4.9 --- Limiting factors for BaP degradation --- p.150 / Chapter 4.10 --- Further Investigations --- p.152 / Chapter 5. --- Conclusion --- p.155 / Chapter 6. --- References --- p.158

Plantios mistos de eucalipto com leguminosas arbóreas vêm sendo estabelecidos a fim de aumentar os teores de nutriente no solo, e estimular a ciclagem biogeoquímica. Para tanto, este trabalho teve como objetivo avaliar alguns atributos microbianos e químicos do solo e serapilheira dos primeiros 20 meses de crescimento de uma segunda rotação de plantios puros e mistos de E. grandis e A. mangium. Foram coletadas amostras de solo e raízes (profundidade de 0-10 cm) e serapilheira aos 2, 7, 14 e 20 meses após o plantio dos seguintes tratamentos: monoculturas de A. mangium e E. grandis sem fertilização nitrogenada, monocultura de E. grandis fertilizado com N e plantio misto entre E. grandis e A. mangium (50E:50A). Dentre os atributos microbianos e bioquímicos do solo foram avaliados o C e N da biomassa microbiana, respiração basal, a atividade das enzimas desidrogenase, celulase, urease, fosfatase ácida, fosfatase alcalina, número de esporos e taxa de colonização de fungos micorrízicos arbusculares. Os atributos químicos do solo avaliados foram: C e N total do solo, P disponível, pH e umidade. Na serapilheira foram avaliadas as concentrações de C, N e P total. As informações obtidas revelaram que houve um efeito cumulativo recorrente da primeira rotação, principalmente na área plantada com A. mangium. Foi possível verificar que no inicio dos plantios florestais os teores de C e N total e P disponível no solo não se alteram drasticamente. Em contrapartida, os conteúdos totais desses elementos na serapilheira são extremamente dinâmicos, dissimilares entre os tratamentos e altamente relacionados com os atributos microbianos e bioquímicos do solo. A. mangium apresentou uma serapilheira com menor relação C/N e C/P, e essa característica se manteve quando ela foi planta com E. grandis, que também apresentou diminuição da relação C/N e C/P da serapilheira. Isso pode ter ocorrido em função ao estimulo da microbiota do solo, sobretudo das fosfatases e pela associação com das plantas com fungos micorrízicos arbusculares. Neste caso, presume-se que a A. mangium é a grande responsável por favorecer a maior atividade microbiana e colonização de FMA em E. grandis no plantio misto quando comparado com os plantios puros de E. grandis. As fosfatases ácida e alcalina foram os atributos microbianos que mais se alteraram com o desenvolvimento vegetal, mostrando a sensibilidade dessa variável aos sistemas de plantio. Estes resultados são informações importantes não apenas sobre a capacidade do plantio misto de E. grandis e A. mangium em maximizar a ciclagem de N e P, mas também sobre o conhecimento da dinâmica desses elementos e da microbiota do solo no inicio dos plantios florestais, fato este pouco abordado pela literatura. / Intercropped plantation between leguminous and eucalyptus has been establishment to increase soil nutrients content and stimulate the biogeochemistry cycling. This work aim evaluated some microbial and chemistry attributes of soil and litter in the early stages of the second rotation of mixed and pure plantations of E. grandis and A. mangium. Soil and roots samples (at 0-10 cm depth) and litter samples were collected at two, seven, 14, and 20 months after planting in the following treatments: monocultures of A. mangium and E. grandis, a monoculture of E. grandis with N-fertilizer, and an intercropped plantation with E. grandis and A. mangium (50E:50A). The intercropped plantation area was subdivided and soil samples collected around of the A. mangium and E. grandis individuals in order to compare the rhizosphere effects of each tree species within this treatment. The microbial and biochemistry attributes measured were: microbial biomass C and N, microbial respiration, metabolic quotient (qCO2), metabolic microbial, enzymatic activity (dehydrogenase, cellulase, urease, acid phosphatase and alkali phosphatase), spore diversity and density and root colonization by arbuscular mycorrhizal fungi. The chemistry soil attributes evaluated were: Total C, total N, available P, pH and moisture. Total C, N and P litter were evaluated. There was a cumulative effect due the first rotation, mainly at A. mangium monoculture. Soil chemistry attributes (total C, total N and available P) were little altered in early stage of the plantations. However, litter attributes were extremely dynamics, dissimilar between treatments and correlated with the soil microbial attributes. A. mangium showed decrease of the litter C/N and C/P ratios (pure and intercropped plantation). In this case, E. grandis also decrease the litter C/N and C/P ratios when planted with A. mangium. Intercropped plantation stimulated the microbial activity, mainly the phosphatases and root colonization by arbuscular mycorrhizal fungi. Thus, presumably A. mangium is the greater responsible to stimulate the soil microbial community in intercropped plantation, favoring E. grandis. Alkali and acid phosphatase were microbial attributes more drastically altered with plant development, showing the sensibility these variables. These results are important information not only about to intercropped plantation capacity for maximize N and P cycling, but also about the knowledge of the dynamic of these elements and the soil microbial in early stages of the forestry plantations, which is little reported to literature.

Submitted by Liliane Ferreira (ljuvencia30@gmail.com) on 2018-04-02T14:29:46Z No. of bitstreams: 2 Dissertação - Marcus Vinícius Forzani Araújo - 2017.pdf: 1549598 bytes, checksum: 0807ed298bd63e9574018c8c399c3ca5 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-04-02T14:41:18Z (GMT) No. of bitstreams: 2 Dissertação - Marcus Vinícius Forzani Araújo - 2017.pdf: 1549598 bytes, checksum: 0807ed298bd63e9574018c8c399c3ca5 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-04-02T14:41:18Z (GMT). No. of bitstreams: 2 Dissertação - Marcus Vinícius Forzani Araújo - 2017.pdf: 1549598 bytes, checksum: 0807ed298bd63e9574018c8c399c3ca5 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-10-13 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Characterized as extremely important, the soil is a complex environment and it shelters a great diversity of microorganisms. However, little is known about the diversity and ecology of the soil microbiota. Thus, the first part of this dissertation reviews the methodological evolution used to characterize the diversity and abundance of microorganisms found in soil. The second part consists of the application of two methodologies reviewed in the previous chapter, serial dilution and solid medium plating, to estimate free-living nitrogen fixing microorganisms, and fumigation-extraction to estimate soil microbial biomass (BMS). The last part employs the most modern microbial soil characterization technique, the metagenomics of 16S rRNA. Hence, our initial hypothesis was that sugarcane fields’ soils would have better soil microbiological indicators than grasslands’ soils. The results confirmed that the hypothesis was partially correct, and it was possible to find about 140% more free-living diazotrophic colony-forming units (CFUs) and a 17% richer alpha diversity in sugarcane fields’ soils than in grasslands’ soils. The beta diversity between sugarcane plantations and pastures presented clear differences. However, sugarcane fields’ soils obtained about 25% less BMS than grasslands’ soils. In relation to the bacterial phyla, the grasslands have more Actinobacteria, Chloroflexi and Planctomycetes and sugarcane fields have a greater number of TM7 and bacteria that were not identified, being Proteobacteria and Acidobacteria the dominating phyla in both types of soil. Although the results of nitrogen fixers and microbial biomass appear to be conflicting, it is an indication that the diazotrophic community undergoes with a diverse biotic and abiotic influences than the total community of soil microorganisms, and thus respond differently. / Caracterizado como de extrema importância, o solo é um ambiente complexo e que abriga uma grande diversidade de micro-organismos. Entretanto ainda pouco se sabe sobre a diversidade e ecologia da microbiota do solo. Deste modo, a primeira parte desta dissertação revisa a evolução metodológica empregada para caracterizar a diversidade e abundância dos micro-organismos encontrados no solo. A segunda parte consiste na aplicação de duas metodologias revisadas no capítulo anterior, a de diluição seriada e plaqueamento em meio sólido, para estimar micro-organismos fixadores de nitrogênio de vida-livre, e a fumigação-extração, para estimar a biomassa microbiana do solo (BMS). E a última parte emprega a técnica mais moderna de caracterização das comunidades microbianas de solo, a técnica de metagenômica de 16S rRNA. À vista disso, a nossa hipótese inicial era que solos de canavial teriam indicadores microbiológicos de solo melhores do que solos de pastagem. Os resultados comprovaram que a hipótese estava parcialmente correta, sendo possível encontrar cerca de 140% a mais de Unidades Formadoras de Colônias (UFCs) de diazotróficos de vida-livre e uma diversidade alfa 17% mais rica em solos de canaviais do que em solos de pastagens. A diversidade beta entre canaviais e pastagens apresentou diferenças nítidas. Entretanto, os solos de canaviais obtiveram cerca de 25% a menos de biomassa microbiana do solo do que solos de pastagens. Em relação aos filos bacterianos, os pastos possuem mais Actinobacteria, Chloroflexi e Planctomycetes e canaviais possuem maior número de TM7 e bactérias que não foram identificados, sendo Proteobacteria e Acidobacteria os filos dominantes nos dois tipos de solo. Apesar de parecerem conflitantes os resultados de fixadores de nitrogênio e biomassa microbiana, é um indicativo de que a comunidade de diazotróficos sofrem influências bióticas e abióticas diversas do que a comunidade total de micro-organismos do solo, e desta forma, respondem de forma diferente.

A cana-de-açúcar é atualmente a cultura de maior importância agrícola do Estado de São Paulo, a partir da qual são gerados açúcar e etanol, além de vários outros subprodutos. No entanto, com a expansão das fronteiras agrícolas e alterações nas práticas de manejo, ocorre atualmente um momento de adequação de tal cultivo, que visa uma maior produtividade e sustentabilidade de produção. Para isto, dentre outros fatores, o papel da comunidade microbiana presente nos solos pode ter fundamental importância, auxiliando no melhor desenvolvimento da planta. No entanto, pouco se sabe sobre a comunidade microbiana existente nos solos cultivados com cana-de-açúcar. Dessa forma, este trabalho teve como objetivo avaliar a diversidade e a abundância de fungos em solos de cultivo de cana-deaçúcar no estado de São Paulo, em áreas sob diferentes atributos químicos, físicos e de manejo. Objetivou-se também, verificar a ocorrência de padrões biogeográficos na estruturação de tais comunidades. Para isso, foi realizada a análise da estrutura das comunidades fúngicas por polimorfismo de comprimento de fragmentos de restrição terminal (T-RFLP), juntamente com a quantificação destas comunidades por meio da PCR em tempo real (qPCR) em 476 amostras de solo, obtidas de 11 áreas de cultivo (usinas). Dentro deste conjunto de dados, temos que os atributos químicos, físicos e manejo explicam maiores valores de variância dentro de cada área amostra, mas pouco explicam da variância geral dos dados, sugerindo a ocorrência de padrões biogeográficos das comunidades de fungos neste ambiente. Tal ocorrência foi confirmada pela significância estatística da correlação entre distância e dissimilaridade das comunidades de fungos, dando suporte a geração dos primeiros mapas biogeográficos de fungos em tais solos. Adicionalmente, a abundância de fungos mostrou-se relacionada com a produtividade da cultura, indicando este ser um dos fatores que modulam a produtividade de cana-de-açúcar nas áreas avaliadas. / The sugarcane is nowadays, the most important crop in the State of São Paulo, serving as the raw material for the production of sugar and ethanol, besides many others by-products. Considering the expansion of agricultural barriers, and shifts in fields management, such cultivation is under a re-arrangement process, aiming to a higher productivity and sustainability. In order to achieve that, among other factors, the role of microbial communities present in soils can be essential to support plant development. However, a few is known about the microbial community under sugarcane crop production soils. Hence, this work intended to evaluate the fungi diversity and abundance in soils cultivated with sugarcane in the State of São Paulo, exploring areas under distinct chemical and physical attributes and also distinct management practices. It was also aimed to determine the occurrence of biogeographically patterns in the structure of such communities. Indeed, it was made the analysis of the fungal community structure by terminal restriction fragment length polymorphism (T-RFLP), together with the quantification of these communities by real time PCR (qPCR) in 476 soils samples, collected in 11 areas cultivated with sugarcane (mills). Within this dataset, it was found that chemical, physical and management attributes explain higher values of variance within each sampled area, but explain little about the total variance of data, suggesting the occurrence of biogeographically patterns in fungal communities in this environment. It was confirmed by the statistical significance of the correlation between distance and dissimilarity of fungal communities, supporting the generation of very first biogeographically maps in such soils. Additionally, the abundance of fungi revealed to be related with sugarcane productivity, indicating this issue as one of the factors modulating the sugarcane productivity in the evaluated areas.

Avaliou-se o efeito das doses dos metais, cádmio e níquel, na velocidade de degradação da matéria orgânica adicionada ao solo através de torta de filtro em ensaios de respirometria. O solo utilizado, Latossolo Vermelho Amarelo de textura média, foi amostrado a uma profundidade de 0-20 cm. Foram aplicadas doses de 0; 28; 56; 112 e 200 mg kg-1 de cádmio e 0; 62,5; 125; 250 e 500 mg kg-1 de níquel como CdCl2.2,5H2O e NiCl2.6H2O, respectivamente. As doses de torta de filtro foram de 0, 40, 80 e 120 Mg ha-1, definidas com base nas doses normalmente aplicadas no cultivo de cana. Na instalação dos experimentos, as massas requeridas dos metais foram adicionadas a 800 g de solo, após a incorporação da torta de filtro. Foram empregados potes de vidro com tampa vedante e capacidade de 2 L, contendo solo umedecido para atingir 80% da capacidade máxima de retenção de água. A incubação foi conduzida à temperatura de 28oC durante 72 dias. Os dois ensaios de incubação, um para cádmio e outro para níquel, foram conduzidos em esquema fatorial 5 x 4, em delineamento inteiramente casualizado, com 3 repetições. O CO2 liberado do solo foi absorvido em solução de NaOH e quantificado por método condutimétrico. No solo, após o período de incubação, foram determinados: pH (H2O) e a fração do metal extraído por solução DTPA-TEA pH 7,3. Curvas de respiração foram estabelecidas a partir das quantidades acumuladas de CO2 liberado e ajustadas ao modelo duplo de cinética de primeira ordem. O efeito dos metais foi avaliado em função das constantes de velocidade obtidas nos ajustes. Considerando-se o pH do solo, medido no final da incubação, observou-se tendência de elevação do pH em função das doses de cádmio aplicadas nas maiores doses de torta de filtro. Para o níquel, na ausência de torta de filtro ou na dose de 40 Mg ha-1 não ocorreu resposta, por outro lado, observou-se um efeito mais expressivo a elevação do pH na dose de 120 Mg ha-1 de torta de filtro e na presença de 500 mg kg-1 de níquel. De maneira geral, a extração de cádmio do solo, ao final do experimento, através da solução DTPA-TEA pH 7,3 foi maior que a de níquel. A torta de filtro afetou mais a eficiência de extração do níquel do que do cádmio. A produção total de CO2 acumulada foi afetada negativamente tanto pelo cádmio como pelo níquel em proporção direta às doses dos metais. Observou-se ainda um efeito positivo dos metais sobre a velocidade de degradação, ou seja, os parâmetros cinéticos indicaram que, sob efeito dos metais estudados, a torta de filtro é degradada em menor proporção, mas de maneira mais rápida. Considerando-se a porcentagem de degradação da matéria orgânica, conclui-se que o efeito negativo de cádmio e níquel estudados foi similar, com uma redução de 46 e 48 %, respectivamente. O efeito do cádmio na degradação foi mais pronunciado na dose de 50 mg kg-1, enquanto que o efeito do níquel variou linearmente com o aumento das doses do metal. / It was evaluated the effect of doses of nickel and cadmium on the degradation of the organic matter applied to soil by means of filter cake in a respirometry trial. Soil employed was Latossolo Vermelho Amarelo, sampled in the field at depth of 0-20 cm. It were applied doses of 0; 28; 56; 112 and 200 mg kg-1 Cd and 0; 62,5; 125; 250 and 500 mg kg-1 Ni as CdCl2.2,5H2O and NiCl2.6H2O, respectively. The filter cake doses were 0, 40, 80 e 120 Mg ha-1, defined in a sugarcane crop basis. In the start of the experiment, a require metals mass was added to 800g soil mass, after filter cake incorporation. It were applied 2 liter glass flask with a closed stopple. Soil moisture was adjusted to 80% of the water holding capacity value and the incubation carried out at 28 degrees Celsius during 72 days. The two trials, one for nickel and the other for cadmium, were established in a 5 x 4 factorial and fully randomized design with 3 replications. The CO2 gas evolved from soil due to microbial respiration was absorbed in a fixed volume of NaOH solution and its amount was then estimated by a condutometric method. After the incubation period the soil pH and the metal content extracted by the DTPA-TEA pH 7,3 solution were determined. Respiratory curves were established through the accumulated amount of CO2 evolved from soil using a double phase first order kinetics model and the metal effect was analyzed with basis on the first order rate constants. The metals effect was evaluated in constant velocity role obtained by adjusts. Considering the soil pH, measured at the end of the incubation period, it was observed a trend of pH increasing as doses of cadmium increase, applied on the greater filter cake doses. Concerning nickel, in filter cake absence or in the 40 Mg ha-1 dose of there was no response and the most expressive effect of pH rising occurred in dose of 120 Mg ha-1 of filter cake in the presence of 500 mg kg-1 nickel. In general the extraction of cadmium in the soil, at the end of the experiment, by solution DTPA-TEA pH 7,3 was greater than nickel. The filter cake affected more the efficiency extraction of nickel than cadmium. The accumulated total CO2 production was affected negatively as cadmium as for nickel in direct ratio to the doses of the metals. It was still observed a positive effect of metals on the degradation speed of filter cake organic matter. The kinetic parameters indicated that, under effect of studied metals, the filter cake is degraded in a lesser ratio, but in a faster manner. Considering the percentage of organic material degradation, it was concluded that the negative effect of cadmium and nickel was similar, with a reduction of 46 and 48 %, respectively. The cadmium effect on the degradation was more pronounced with 50 mg kg-1, while the nickel effect rose linearly with metal doses increase.