Dissertations / Theses on the topic 'Mushroom compost'

Spent mushroom compost (SMC) is an agricultural waste disposed of in an unsustainable and environmentally degrading manner - mainly in landfills. For every 1 kg of mushrooms grown, approximately 5 kg of SMC is produced, where current generation in the UK is 200,000 tJa. Coal tailings, an industrial by-product from coal mining, are also discarded untenably in lagoons; removing these deposits will eliminate the associated environmental hazards. This project aimed to combine these waste materials into a suitable 'green' fuel for industry through thermal treatment, to produce energy from a sustainable source. Not only will this alleviate the issues regarding existing waste management strategies, but will also attempt to mitigate the environmental impacts of energy generation from nonrenewable sources, such as anthropogenic climate change, through the generation of renewable energy. This PhD research has shown that both materials had high moisture contents, which negatively impacted the calorific value (CV). Drying, though expensive, would thus be required prior to pelletisation and thermal treatment. Key pelletisation parameters were identified and manipulated to control product quality. Optimal values were experimentally-determined for pellet composition (50:50 wt% SMC:coal tailing ratio), moisture (10.5 %) and pressure (6000 psi/41 MPa); such pellets had a NCV of 16.11 MJ/kg. As these pellets were still friable, additional studies were carried out to further improve pellet quality, in tenns of density, tensile strength and durability. Elevated temperatures and steam were considered, in addition to the use of starch and caustic soda binders, which were all successful to varying degrees. Combustion, gasification and pyrolysis tests compared the raw SMC to SMC-coal tailing pellets, where pellet combustion performed better than the SMC alone, and fluidised-bed combustion was more efficient than the packed-bed. Although pyrolysis worked well, the CV of the fuel products were low, whereas gasification was unsuccessful. Consequently, in-depth studies into pellet combustion in a laboratory scale fluidised-bed were perfonned, examining: (i) combustionl fluidisation air flowrates (4.9-10.7 kg/hr); (ii) fuel pellet feedrates (2.02-4.58 kg/hr); and (iii) sand bed depths (0.22-0.30 m). The impacts on temperatures, combustion efficiency and gas concentrations, including acid gas species were analysed. The most favourable operating conditions resulted in high temperatures for efficient energy recovery, with minimal pollutants, although the addition of secondary air jets could further improve the already high combustion efficiencies. While gaseous pollutants are unlikely to be an issue, as the emissions produced generally conformed to the Waste Incineration Directive, efficient particulate collection will be required to remove flyash from the gas stream prior to release to the atmosphere. Industrial implications were explored for heat and power generation, where mass and energy balances for a theoretical furnace, boiler and turbine set-up were completed for various fuel throughputs. Excess heat from the process could be utilized to dry the initial materials, but the economic analysis showed this would be costly - totalling 7) of overall pelletisation expenses. Assuming an overall process efficiency of 18.6 %, a steam turbine could generate over 10 MWe, based on an SMC-coal tailing pellet feedrate of 400,000 tJa - to simulate a large, centralised energy-from-waste facility. FLUENT, a mathematical model, was able to effectively replicate the results of the experimentation and was then used to model particle elutriation and entrainment to assess the suitability of the transport disengagement height provided.

In this research spent mushroom compost ash (SMCA), the incinerated product of spent mushroom compost (SMC), was investigated for use in Portland cement (PC) based materials. Characterisation studies found that SMCA contained, among other things, calcium and alkali sulphates. The sulphate content within SMCA was utilised in this investigation as: (A) a set retarder, replacing calcium sulphate in PC and (B) a sulphate activator of pulverised fuel ash (PF A) in PF AlPC blends. Results demonstrated that SMCA was effective at both retarding set in PC and chemically activating PF A in PF AlPC blends. Moreover, in both applications, mineralogical changes and compressive strength results during the fIrst 24 hours of hydration indicated that the presence of SMCA in PC accelerated the early hydration and strength development (lday) compared to PC without SMCA. However, the long-term strength (at 90 days) was detrimentally affected when SMCA was present within either PC or PCfPF A blends. Although the usefulness of SMCA in cement based materials has been demonstrated other factors, such as the presence of alkalis and chlorides in the pore solution, may limit its use and as a result further investigation is required.

This study aimed at seeking for the solution to recover the energy from agriculture waste in the Mekong Delta, Vietnam. The spent mushroom compost - a residue from the mushroom growing - was chosen for co-digestion with pig manure in anaerobic batch and semi-continuous experiments. The results showed that in case of spent mushroom compost made up 75% of the mixed substrate, the gained biogas volume was not significantly different compared to the treatment fed solely with 100% pig manure. The average produced biogas was 4.1 L×day-1 in the experimental conditions. The semi-continuous experiments remained in good operation up to the 90th day of the fermentation without any special agitating method application. The methane contents in both experiments were around 60%, which was significantly suitable for energy purposes. These results confirm that spent mushroom compost is possibly an acceptable material for energy recovery in the anaerobic fermentation process
Nghiên cứu này nhằm tìm kiếm giải pháp thu hồi năng lượng từ chất thải nông nghiệp tại ĐBSCL, Việt Nam. Rơm sau ủ nấm - phế phẩm sau khi trồng nấm rơm - được chọn để ủ kết hợp với phân heo trong các bộ ủ yếm khí theo mẻ và bán liên tục. Kết quả cho thấy nếu phối trộn đến 75% rơm sau ủ nấm trong nguyên liệu ủ, tổng lượng khí thu được không khác biệt đáng kể so với thí nghiệm ủ 100% phân heo. Trong điều kiện thí nghiệm, lượng khí thu được trung bình là 4.1 L.ngày-1. Thí nghiệm ủ bán liên tục vẫn vận hành tốt ở ngày thứ 90 mặc dù mẻ ủ không được khuấy đảo. Hàm lượng khí mê-tan đo được chiếm khoảng 60% hoàn toàn có thể sử dụng cho các nhu cầu về năng lượng. Những kết quả thí nghiệm khẳng định có thể sử dụng rơm sau ủ nấm để thu hồi năng lượng thông qua quá trình ủ yếm khí kết hợp

Overuse of the chemical compounds and toxic elements leads to problems and transmission of contaminants and pollutants to humans and other living organisms. One of the industries’ byproducts of the agriculture sector is production of various composts from the organic raw materials that the best type of which is so – called Vermicompost. In this study, effects of raw materials on qualitative and quantitative characteristics of Vermicompost are discussed. To do so, sheep manure, pomegranate peels, spent mushroom compost either singly or double, triple or fourfold chopped corn, sugar beet pulp and sawdust were used. This research project was conducted in a completely randomized design experiment with 23 treatments with 3 replications. Results revealed that various bed combinations exert different effects on Vermicompost quality such that, the Vermicomposting process led to a significant decrease in electrical conductivity (EC) and a significant increase in pH in most of the culture (seed) beds. Also, the levels of Nitrogen, Phosphorous and Potassium in most treatments increased following completion of the vermicomposting process. As a result, this process can be introduced as an organic fertilizer with complete nutrients for improving chemical characteristics of agricultural wastes to usable fertilizers.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
A produção de composto de qualidade para Agaricus bisporus e a pesquisa por linhagens produtivas são alguns dos principais fatores relacionados à produtividades elevadas. Desta forma, foram realizados dois experimentos: 1. a campo, avaliou-se o efeito da suplementação nitrogenada na formulação de dois tipos de compostos, clássico e sintético, para o cultivo de cinco linhagens de A. bisporus: ABI-05/03, ABI-04/02, ABI-06/05, ABI-09/10 e ABI-09/11; 2. avaliou-se a influência de cinco linhagens de A. bisporus no desenvolvimento micelial em dois meios de cultura sólidos (CA, composto ágar; e BDA, batata dextrose ágar). No experimento 1, constatou-se durante o processo de compostagem, pasteurização e condicionamento o composto clássico obteve temperatura média e perda de massa 10,56 e 13,29% superiores ao composto sintético, respectivamente. O composto clássico obteve as maiores eficiências biológicas ao final de 25 dias de produção, pelas linhagens ABI-05/03, ABI-06/05 e ABI-04/02 com valores de 83,95, 79,45 e 77,49%, respectivamente. Além da eficiência biológica, houve uma tendencia de maior produtividade, número e massa de fresca de basidiomas quando as linhagens foram cultivadas em composto clássico. No experimento II as maiores velocidades de desenvolvimento micelial das linhagens de A. bisporus foram observadas nos meios de cultura CA. Concluiu-se que não houve ligação entre os resultados observados nos experimentos I e II em relação ao potencial genético das...
The production of quality compost for Agaricus bisporus and the research for high productivity strains are some important factors involving high yields. Were carried out two expiriments: 1. at field, the effect of the type of nitrogen supplementation was evaluated, elaborating two types of compost, classic and synthetic, cultivating five strains of A. bisporus ABI-05/03, ABI-04/02, ABI-06/05, ABI-09/10 e ABI-09/11; 2. was evaluated the influence of five A. bisporus strains on the rate of micelial growth in different type of culture media (MC, compost media; BDA, potato-dextrose-agar). In the first experiment, the data showed that during the composting process, pasteurization and conditioning, the averages temperatures and weight loss 10,56 and 13,29% higher in the classic compost than the synthetic compost . The classic compost had the higher biological efficiency in the end of the crop (25 days), for the strains ABI-05/03, ABI-06/05 e ABI-04/02 with values of 83,95, 79,45 e 77,49, respectively. Moreover, there was a tendency for higher yields, number and fresh weight of mushrooms when the strains were cultivated in the classic compost. In the second experiment the highest micelial growth rate by the A. bisporus strains were observed in the compost agar media. It was observed that were no relation between the data in experiments I and II, by the genetic potential of the strains

Orientador: Marli Teixeira de Almeida Minhoni
Banca: Meire Cristina Nogueira de Andrade
Banca: Eduardo Bagagli
Resumo: A produção de composto de qualidade para Agaricus bisporus e a pesquisa por linhagens produtivas são alguns dos principais fatores relacionados à produtividades elevadas. Desta forma, foram realizados dois experimentos: 1. a campo, avaliou-se o efeito da suplementação nitrogenada na formulação de dois tipos de compostos, clássico e sintético, para o cultivo de cinco linhagens de A. bisporus: ABI-05/03, ABI-04/02, ABI-06/05, ABI-09/10 e ABI-09/11; 2. avaliou-se a influência de cinco linhagens de A. bisporus no desenvolvimento micelial em dois meios de cultura sólidos (CA, composto ágar; e BDA, batata dextrose ágar). No experimento 1, constatou-se durante o processo de compostagem, pasteurização e condicionamento o composto clássico obteve temperatura média e perda de massa 10,56 e 13,29% superiores ao composto sintético, respectivamente. O composto clássico obteve as maiores eficiências biológicas ao final de 25 dias de produção, pelas linhagens ABI-05/03, ABI-06/05 e ABI-04/02 com valores de 83,95, 79,45 e 77,49%, respectivamente. Além da eficiência biológica, houve uma tendencia de maior produtividade, número e massa de fresca de basidiomas quando as linhagens foram cultivadas em composto clássico. No experimento II as maiores velocidades de desenvolvimento micelial das linhagens de A. bisporus foram observadas nos meios de cultura CA. Concluiu-se que não houve ligação entre os resultados observados nos experimentos I e II em relação ao potencial genético das...
Abstract: The production of quality compost for Agaricus bisporus and the research for high productivity strains are some important factors involving high yields. Were carried out two expiriments: 1. at field, the effect of the type of nitrogen supplementation was evaluated, elaborating two types of compost, classic and synthetic, cultivating five strains of A. bisporus ABI-05/03, ABI-04/02, ABI-06/05, ABI-09/10 e ABI-09/11; 2. was evaluated the influence of five A. bisporus strains on the rate of micelial growth in different type of culture media (MC, compost media; BDA, potato-dextrose-agar). In the first experiment, the data showed that during the composting process, pasteurization and conditioning, the averages temperatures and weight loss 10,56 and 13,29% higher in the classic compost than the synthetic compost . The classic compost had the higher biological efficiency in the end of the crop (25 days), for the strains ABI-05/03, ABI-06/05 e ABI-04/02 with values of 83,95, 79,45 e 77,49, respectively. Moreover, there was a tendency for higher yields, number and fresh weight of mushrooms when the strains were cultivated in the classic compost. In the second experiment the highest micelial growth rate by the A. bisporus strains were observed in the compost agar media. It was observed that were no relation between the data in experiments I and II, by the genetic potential of the strains
Mestre

Orientador Meire Cristina Nogueira de Andrade
Banca: Eduardo Bagagli
Banca: Adriana Zanin Kronka
Banca: Edson Luiz Furtado
Banca: Geysiany Maria de Queiroz-Fernandes
Resumo: Entre os cogumelos cultivados comercialmente, destacam-se o Agaricus bisporus e Pleurotus ostreatus (cogumelo ostra). A preparação do substrato de cultivo é a etapa mais crítica da produção, determinante para a produtividade e custos de produção. O método de preparo do substrato mais utilizado pela indústria de cogumelos é a compostagem curta, seguida de pasteurização e condicionamento. Como ponto chave da compostagem, a comunidade microbiana tem papel fundamental no processo, conferindo seletividade final do substrato de cultivo e, consequentemente, a produtividade. Desta forma, avaliou-se, o manejo da compostagem (tempo de compostagem e temperatura de pasteurização) e o efeito sob a comunidade microbiana, composição final do substrato e produtividade de cogumelos. Para isto, foram realizados três experimentos. Nos experimentos 1 e 2, avaliou-se a influência do tempo de compostagem (fases I e II), a comunidade microbiana (bactérias e fungos), variáveis físico-químicas, produtividade, número de cachos e precocidade do cogumelo P. ostreatus. Os resultados obtidos mostraram que o manejo da compostagem influenciou a comunidade microbiana e, consequentemente, a seletividade final do substrato de cultivo, com reflexos na produtividade e número de cachos. A comunidade microbiana apresentou menor diversidade na compostagem com menor duração e, menor perda de carbono, nitrogênio e açúcares solúveis, resultando em substrato mais seletivo ao cogumelo P. ostreatus, demostrando melhores resultados de produtividade. No experimento 3, avaliaram-se, a comunidade microbiana e indicadores químicos do composto final, em diferentes situações de pasteurização durante a preparação do substrato para o cultivo do cogumelo A. bisporus. Para isto, foram simuladas em biorreatores três diferentes situações de pasteurização (57°C por 6 horas, 60°C por 2 horas e 68°C por 2 horas). ...
Abstract: Among many commercial mushrooms cultivated, Agaricus bisporus and Pleurotus ostreatus (oyster mushroom) are the most important for mushroom industry. The cultivation substrate preparation is the most critical step in mushroom production, determining productivity and production costs. The substrate preparation method most used by mushroom industry is short composting, followed by pasteurization and conditioning. As key point of composting, microbial community plays an important role during the process, determining final substrate selectivity and mushroom yield. Thus, aimed evaluate the influence of composting management (composting time and pasteurization temperature) in microbial community, final substrate composition and mushroom yield. For this, were conducted three experiments. In experiments 1 and 2 were evaluated the influence of composting time (phases I and II), microbial community (bacteria and fungi), physicochemical variables and P. ostreatus yield. The results showed that composting management (time of phases I and II) influenced the microbial community, substrate selectivity and consequently mushroom yield and number of cluster. The microbial community showed less diversity in composting with shorter duration, as well as the losses of carbon, nitrogen and soluble sugars, showing higher selectivity of substrate for P. ostreatus, as observed in yield results. In experiment 3, were evaluated the microbial community (bacteria and fungi) and some physiochemical variables in different pasteurization conditions during substrate preparation for A. bisporus cultivation. Were simulated in bioreactors three different pasteurization conditions (57°C for 6 hours, 60°C for 2 hours and 68°C for 2 hours). The sequencing of microbial community revealed the predominance of phyla Proteobacteria and Firmicutes during phase I and II composting, respectively. The most abundant bacterial genera were ...
Doutor

Orientadores: Helena Teixeira Godoy, Adriana Dillenburg Meinhart
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos
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Resumo: Dentre as propriedades funcionais apresentadas pelos cogumelos, destacam-se as propriedades antioxidantes. A principal classe de compostos que colaboram com a atividade antioxidante são os compostos fenólicos. Alguns trabalhos já relataram a presença de compostos fenólicos em diversas espécies de cogumelos, porém estudos que avaliam o perfil dos compostos fenólicos em cogumelos comestíveis cultivados no Brasil são muito escassos. O objetivo desse trabalho foi identificar e quantificar os compostos fenólicos presentes em 4 espécies de cogumelos comestíveis (Agaricus bisporus, Pleurotus ostreatus, Pleurotus ostreatoroseus e Lentinula edodes) produzidas no Brasil, portanto foi desenvolvida uma metodologia de análise por eletroforese capilar através da otimização univariada do pH e análise da curva de mobilidade dos compostos fenólicos. Foi possível separar 11 compostos fenólicos já relatados em cogumelos. O método utilizou os seguintes parâmetros: 175 mmol.L-1 de ácido bórico, pH 8,5, 25°C, 30 kV, capilar 50 µm x 72 cm injeção de 50 mbar durante 5 segundos e detecção em 210 nm, em um tempo de corrida de 21 minutos. Foi realizado um estudo multivariado, utilizando-se um planejamento composto central (otimizando a concentração de ácido clorídrico, tempo e temperatura de extração) para a otimização da extração e hidrólise ácida dos compostos fenólicos presentes na matriz. A condição ótima de extração e hidrólise foi obtida analisando-se 0,3 g de amostra, com 6 mL de ácido clorídrico a 2 mol.L-1, sob refluxo a 79,9°C por 30 minutos. Dentre os compostos investigados, foram identificados e quantificados os ácidos homogentísico, p-cumárico e cinâmico, com predominância do último. O teor de ácido homogentísico variou entre 169,85 e 388,6, p-coumárico entre 25,97 e 103,32 e ácido cinâmico entre 23,38 e 454,07, todos em mg.kg-1 de amostra liofilizada. Foi possível visualizar diferença no perfil dos compostos fenólicos de cogumelos de diferentes variedades. O tipo de substrato no qual os cogumelos são cultivados também foi significativo para os resultados. Da mesma forma, foi possível identificar diferenças em amostras com diferentes graus de maturação
Abstract: Among the celebrated functional properties of mushrooms, it calls the attention their antioxidant properties. Antioxidant compounds of the mushrooms are mailing the phenolic compounds. Some studies have reported the presence of phenolic compounds in several species of mushrooms, but studies that evaluate the phenolic content in edible mushrooms grown in Brazil are scarce. The aim of this study was to identify and measure phenolic compounds present in four species of edible mushrooms (Agaricus bisporus, Pleurotus ostreatus, Lentinula edodes and Pleurotus ostreatoroseus) cultivated in Brazil. A methodology of analysis was developed using capillary electrophoresis with pH univariate optimization and analysis of the mobility curve of phenolic compounds, separating 11 phenolic compounds already described in the literature. The following conditions were determinated: 175 mmol.L-1 of boric acid, pH 8.5, 25°C, 30 kV, capillary of 50 µm x 72 with an injection of 50 mbar for 5 seconds and detection at 210nm, at a running time of 21 minutes. A multivariate study was conducted for the optimization of extraction and acid hydrolysis of phenolic compounds present in pattern. It was used a central composite design (optimizing the concentration of hydrochloric acid and extraction time and temperature). It was determined that the optimum condition for extraction and hydrolysis was samples of 0.3 g, 6 mL of hydrochloric acid at 2 mol.l-1 under reflux at 79.9°C for 30 minutes. Among other compounds investigated, homogentisic, p-coumaric and cinnamic acids were identified and quantified, with predominance of the last one. Homogentisic acid content varied from 169.85 to 388.6; p-coumaric acid from 25.97 to 103.32 and cinnamic acid from 23.38 to 454.07, in mg.kg-1 of lyophilized sample. Differences in phenolic content were observed for different varieties of mushrooms, type of substrate on which the mushrooms were grown and stage of maturity
Mestrado
Ciência de Alimentos
Mestre em Ciência de Alimentos

Ganoderma lucidum é um cogumelo medicinal consumido tradicionalmente nos países asiáticos que apresenta diversos efeitos benéficos comprovados. Apesar de todos os estudos sobre seus compostos bioativos, a melhor forma de enriquecimento de meios de cultivo visando aumentar a produção destes compostos ainda se mantém incerta. Além disso, poucos trabalhos tratam dos efeitos sobre o desempenho de animais de produção. Para testar diferentes meios de cultivo de micélio de G. lucidum, foram testados resíduos agrícolas (fermentação em estado sólido) e diferentes açúcares e aminoácidos (em meios de cultura com ágar), para avaliar o crescimento do micélio de G. lucidum. O fornecimento de farinha com micélio de G. lucidum obtido por fermentação em meio sólido (grãos de trigo) para coelhos também foi avaliado. O micélio de G. lucidum desenvolveu-se bem nos resíduos agrícolas, destacando-se a casquinha de soja que apresentou maior taxa de crescimento e também maior concentração de β-glucanas. No experimento em placas, o G. lucidum também se desenvolveu bem, destacando-se os meios que continham celobiose e tirosina, que apesar de apresentarem menores taxas de crescimento e crescimento total em 10 dias, produziram micélios com maior concentração de β-glucanas e capacidade antioxidante em equivalentes trolox (TEAC), respectivamente. Os coelhos não apresentaram sinais de intolerância ao consumo de ração com diferentes concentrações farinha com micélio de G. lucidum. Todos os parâmetros de desempenho e avaliação de carcaça foram adequados à idade em que foram abatidos. A avaliação histológica dos órgãos apresentou alterações nas células renais (tubulares e glomerulares), apontando para uma possível lesão renal conforme aumentou a concentração de farinha com micélio na ração. A avaliação histomorfométrica resultou em maior altura de vilosidade de íleo e menor largura de vilosidade de jejuno na concentração 0,5%, e redução do diâmetro de cripta conforme o aumento da concentração de farinha com micélio. Estes resultados indicam a possibilidade de estudos mais aprofundados sobre os aspectos de utilização de celobiose e tirosina para produção de compostos bioativos, e sobre aspectos de toxicidade do micélio deste cogumelo para segurança no fornecimento para animais.
Ganoderma lucidum is a medicinal mushroom traditionally consumed in Asian countries that presents several beneficial effects already verified. Despite all studies about their bioactive compounds, the best cultivation media enrichment aiming to increase the production of these compounds is still uncertain. Besides, few studies are related to the performance of production animals. In order to test different cultivation media for G. lucidum mycelia, agricultural residues (solid state fermentation) and different sugars and aminoacids (in culture media with agar), were tested to evaluate G. lucidum mycelium growth. Supply of flour with G. lucidum mycelia obtained by solid state fermentation (wheat grain) for rabbits was also evaluated. Mycelium of G. lucidum developed very well in all agricultural residues, soybean hulls was the residue that presented higher growth rate and higher concentration of β-glucans. In the cultivation media experiment, G. lucidum also developed well, media that contained cellobiose and tyrosine, despite presenting lower growth rates and total growth within 10 days, produced mycelia with higher concentration of β-glucans and trolox equivalent antioxidant capacity (TEAC), respectively. Rabbits did not show any sign of intolerance to feed with different concentrations of flour with G. lucidum mycelium. All performance parameters and dressing percentages were adequate to the age at which they were slaughtered. Histological evaluation of organs presented alterations in renal cells (tubular and glomerulus), indicating a possible renal lesion according to the increase of flour with mycelium in feed. Histomorphometric evaluation showed increased vilous height in ileum and decreased vilous width of jejunus at 0.5% concentration, and decrease in crypt diameter according to the increase of concentration of flour with mycelium in feed. These results indicate the possibility of more studies regarding the aspects about cellobiose and tyrosine utilization for the production of bioactive compounds, and about toxicity of this mushroom mycelia, assuring the safety in supplying this product for animails.

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FAPEAM - Fundação de Amparo à Pesquisa do Estado do Amazonas
The increase in microbial resistance to available antibiotics and the continuous decrease observed in the number of the new antimicrobial agents approved by Food and Drug Administration creates a constant search for new compounds. To do so, one of the strategies to be followed consists in exploring understudied natural sources, as often organisms obtained new ecosystems are associated with new chemical diversity. In this context are the basidiomycetes, which have been reported as promising sources of antimicrobial and, despite this potential and great biological diversity, the species found in the Amazon have been insufficiently studied. The objective of this study was to evaluate the antimicrobial activity of eight basidiomycetes isolated from Manaus and Boa Vista. The fungi were grown in two liquid media (malt and GLP) and malt-agar medium, in the latter case were held in two incubation conditions: absence and presence of light. The fungi were maintained at room temperature in all treatments. The tests used the filtered liquid from the growth medium for seven to 63 days, besides extracts with ethyl acetate from growing on solid media (ten days). The pathogens tested were Escherichia coli CBAM 001, Staphylococcus aureus CBAM 324, Candida albicans CFAM 1342, Aspergillus niger CFAM 161 and Penicillium sp. CFAM 059, by agar diffusion method. The filtered with positive activity, were submitted to microdilution test for identification of minimum inhibitory concentration. Molecular identification was performed by amplification and sequencing of the rDNA fragment, with subsequent comparison of the sequences obtained with those deposited in the National Center for Biotechnology Information database. Of filtered tested, seven inhibited the growth of C. albicans CFAM 1342, all of Oudemansiella canarii 1528 cultivation, as medium malt, as in GLP. At medium malt, this basidiomycete produced antifungal compounds at seven and 49 days of cultivation, while in the medium GLP produced from the 21 days of cultivation, until the 49th day. The minimum inhibitory concentration was found for the filtrate obtained after 28 days of GLP cultivation medium (2.5 mg/ml) and the filtrate obtained after 49 days malt medium (5 mg/ml). The extracts obtained with ethyl acetate inhibited C. albicans CFAM 1342, Penicillium sp. CFAM 059, E. coli CBAM 001 and S. aureus CBAM 324. This method was found more satisfactory results, as well as Oudemansiella canarii 1528, six other basidiomycetes (Basidiomycete 347, Pleurotus sp. 474, Gloeophyllum sp. 1153, Trametes sp. 1232, Trametes sp. 1540 e Earliella scabrosa 1552) showed antimicrobial activity. Thus, the results allow to observe the influence of culture conditions for the production of antimicrobial compounds, as well as choice of solvents, further studies are needed to determine the optimum conditions for production and extraction thereof.
O aumento da resistência microbiana aos antibióticos disponíveis e o decréscimo contínuo observado no número de novos antimicrobianos aprovados pela Food and Drug Administration gera uma constante busca por novos compostos. Para tanto, uma das estratégias a ser seguida consiste na exploração de fontes naturais pouco estudadas, pois frequentemente organismos obtidos de novos ecossistemas estão associados à nova diversidade química. Neste contexto destacam-se os basidiomicetos, que vêm sendo relatados como promissoras fontes de antimicrobianos e, apesar deste potencial e da grande diversidade biológica, as espécies ocorrentes na Amazônia têm sido pouco estudadas. Assim, o objetivo deste trabalho foi avaliar a atividade antimicrobiana de oito basidiomicetos isolados de Manaus e Boa Vista. Os fungos foram cultivados em dois meios líquidos (malte e GLP) e em meio ágar-malte, neste último caso foram mantidos em duas condições de incubação: ausência e presença de luz. Em todos os tratamentos os fungos foram mantidos em temperatura ambiente. Os testes utilizaram os filtrados provenientes do crescimento em meio líquido de sete a 63 dias, além de extratos obtidos com acetato de etila a partir do cultivo em meio sólido (dez dias). Os patógenos testados foram Escherichia coli CBAM 001, Staphylococcus aureus CBAM 324, Candida albicans CFAM 1342, Aspergillus niger CFAM 161 e Penicillium sp. CFAM 059, através do método de difusão em ágar. Os filtrados com atividade positiva foram submetidos a teste de microdiluição para identificação da concentração inibitória mínima. Foi realizada a identificação molecular através de amplificação e sequenciamento do fragmento do rDNA, com posterior comparação das sequências obtidas com as depositadas no banco de dados do National Center for Biotechnology Information. Dos filtrados testados, sete inibiram o crescimento de C. albicans CFAM 1342, todos provenientes do cultivo de Oudemansiella canarii 1528, tanto em meio malte, como em meio GLP. Em meio malte, este basidiomiceto produziu compostos antifúngicos aos sete e 49 dias de cultivo, enquanto em meio GLP produziu a partir dos 21 dias de cultivo, até o 49º dia. A concentração inibitória mínima foi identificada para o filtrado obtido aos 28 dias de cultivo em meio GLP (2,5 mg/mL) e o filtrado obtido aos 49 dias em meio malte (5 mg/mL). Os extratos obtidos com acetato de etila além de inibir C. albicans CFAM 1342, inibiram Penicillium sp. CFAM 059, E. coli CBAM 001 e S. aureus CBAM 324. Esta metodologia apresentou resultados mais satisfatórios, pois além de Oudemansiella canarii 1528, outros seis basidiomicetos (Basidiomycete 347, Pleurotus sp. 474, Gloeophyllum sp. 1153, Trametes sp. 1232, Trametes sp. 1540 e Earliella scabrosa 1552) apresentaram atividade antimicrobiana. Deste modo, os resultados obtidos permitiram observar a influência das condições de cultivo para a produção de compostos antimicrobianos, assim como a escolha de solventes, sendo necessários estudos posteriores para determinar a melhor condição para a produção e extração destes compostos.

Orientador: Helena Teixeira Godoy
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos
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Resumo: O aprofundamento do estudo relativo aos compostos naturais com propriedades antioxidantes e antimicrobianas em cogumelos torna-se extremamente relevante tendo em vista a possibilidade de esses compostos serem empregados nas indústrias de alimentos, farmacêutica e de cosméticos. No entanto, não foram encontrados relatos sobre o perfil fenólico de cogumelos comestíveis produzidos no Brasil assim como também não foram encontrados trabalhos que utilizassem quatro ensaios diferentes para avaliar o potencial antioxidante dos cogumelos. Portanto, o objetivo desse trabalho foi avaliar o teor de compostos fenólicos e a capacidade antioxidante e antimicrobiana de 4 espécies de cogumelos comestíveis (A. bisporus, P. ostreatus, P. ostreatoroseus e L. edodes) produzidas no Brasil. Os compostos fenólicos totais (CFT) foram determinados pelo método de Folin-Cioacalteu e a identificação e quantificação individual dos fenólicos foi realizada por cromatografia líquida de alta eficiência (CLAE). A atividade antioxidante foi avaliada por quatro métodos: DPPH (2,2-difenil-1-picrilhidrazila), ABTS+¿ [2,2¿-azino-bis (3-etilbenzo-tiazolina-6-sulfonato)], FRAP (Ferric Reducing Antioxidant Power) e ORAC (Oxygen Radical Absorbance Capacity). O potencial antimicrobiano dos extratos foi avaliado através da concentração mínima inibitória (CIM). Todos os resultados obtidos para as diferentes análises foram analisados levando em consideração o período de colheita, os diferentes processamentos, graus de maturação e substratos de cultivo. No ensaio com o radical DPPH¿ o cogumelo que apresentou maior atividade antioxidante foi o Champignon de Paris (fresco). Já no ensaio com o radical ABTS+¿ e no ORAC os cogumelos que apresentaram maior ação antioxidante foram o Hiratake, o Shimeji (fresco), o Salmon e o Portobello. No ensaio FRAP o Portobello mostrou a maior atividade antioxidante. Os valores médios de CFT variaram entre 0,78 e 10,85 mg EAG/g de amostra liofilizada. Os cogumelos que apresentaram os maiores teores de CFT para o cultivo de verão e de inverno, respectivamente, foram o Hiratake com 10,09 mg EAG/g e 8,45 mg EAG/g, Salmon com 9,61 mg EAG/g e 9,45 mg EAG e Portobello com 9,07 mg EAG/g e 9,60 mg EAG. O processamento empregado ao cogumelo Champignon de Paris diminuiu consideravelmente o teor de CFT (de 7,04 mg EAG/g para 0,90 mg EAG/g - cultivo de verão) enquanto que no processo de secagem empregado ao cogumelo Shimeji não houve diminuição drástica no teor de CFT (de 8,83 mg EAG/g para 7,62 mg EAG/g - cultivo de verão). O grau de maturação influenciou no teor final de CFT mostrando que o Shimeji, colhido em estágio imaturo, possui menor teor de CFT (8,83 mg EAG/g ¿ cultivo de verão e 7,24 mg EAG/g ¿ cultivo de inverno) em comparação ao Hiratake, colhido em estágio mais maturo. Correlações positivas entre os resultados de CFT e os métodos de atividade antioxidante foram encontradas, e essa correlação foi maior para o ensaio com o radical livre ABTS¿+ e o ORAC (r = 0,92 e r = 0,89, respectivamente) do que para o ensaio FRAP (r = 0,45). Já entre o DPPH e os resultados de CFT foi encontrada baixa correlação negativa (r = -0,14). Dentre os compostos investigados por CLAE foram identificados e quantificados apenas os ácidos siríngico, p-cumárico e o trans-cinâmico, com predominância deste último. No ensaio antimicrobiano realizado verificou-se que o crescimento das leveduras C. albicans, C. rugosa, C. tropicalis, C. glabrata e C. utilis foi fortemente inibido pelos extratos aquosos dos cogumelos Shiitake (tora e composto), com CIMs variando entre 0,08 e 0,31 mg/mL. Estes resultados demonstram que os cogumelos comestíveis possuem grande potencial de utilização pela indústria farmacêutica e podem contribuir significativamente para uma boa condição de saúde devido à presença de antioxidantes
Abstract: The study of natural compounds with antioxidant and antimicrobial properties of mushrooms is extremely relevant to the use in the food, pharmaceutical and cosmetic industries. However, no reports were found about the phenolic profile of edible mushrooms produced in Brazil as well as were not found papers describing the use four different tests to evaluate the antioxidant potential of mushrooms. Therefore, the aim of this study is evaluate the phenolic content, antioxidant and antimicrobial activities of four species of edible mushrooms (A. bisporus, P. ostreatus, L. edodes and P. ostreatoroseus) produced in Brazil. The total phenolic compounds (TPC) were determined by Folin-Cioacalteu and the individual identification and quantification of phenolic compounds was performed by high performance liquid chromatography (HPLC). The antioxidant activity was evaluated by four methods: DPPH¿(2,2-diphenyl-1-picrylhydrazyl), ABTS+¿ [2,2 '-azino-bis (3-etilbenzo-thiazoline-6-sulfonate)], FRAP (Ferric Reducing antioxidant Power) and ORAC (Oxygen Radical Absorbance Capacity). The antimicrobial activity of extracts was evaluated by minimum inhibitory concentration (MIC). All the results obtained for the different analyzes were analyzed taking into account the period of harvesting, different processing, maturation degrees and cultivation substrates. In the assay with DPPH, the mushroom with the highest antioxidant activity was A. bisporus white (fresh). In the assays with the radical ABTS+¿ and ORAC, the mushrooms that showed the highest antioxidant activity were P. ostreatus (mature and fresh immature), P. ostreatoroseus and A. bisporus brown. In the FRAP assay the A. bisporus brown. showed the highest antioxidant activity. The average values of TPC ranged varied from 0.78 and 10.85 mg GAE/g lyophilized sample. The mushrooms with highest levels of TPC for the summer and winter cultivation, respectively, were P. ostreatus (mature) with 10.09 mg GAE/g and 8.45 mg GAE/g, P. ostreatoroseus with 9.61 mg GAE/g and 9, 45 mg GAE and A. bisporus brown with 9.07 mg GAE / g and 9.60 mg GAE. The processing used in the A. bisporus white has considerably reduced the content of TFC (from 7.04 mg GAE/g to 0.90 mg GAE/g - summer cultivation) while in the process of drying the mushroom used in the P. ostreatus (immature), no drastic decrease content of the TPC (from 8.83 mg of GAE/g to 7.62 mg GAE/g - summer cultivation). The maturation degree influenced the final content of the TPC showing P. ostreatus, harvested in an immature state, has a lower content of TPC (8.83 mg GAE/g - summer cultivation and 7.24 mg GAE/g - winter cultivation) compared to P. ostreatus, collected in more mature stage. Positive correlations between the results of TPC and methods of antioxidant activity were founded, and this correlation was higher for the test with free radical ABTS+¿ and ORAC (r = 0.92 and r = 0.89, respectively) than for the FRAP test (r = 0.45). Among the DPPH and the results of TFC was found low negative correlation (r = -0.14). Among the compounds investigated by HPLC were identified and quantified only the syringic, p-coumaric and trans-cinnamic acids, with predominance of the latter. In the assay antimicrobial it was found that the growth of yeasts C. albicans, C. rugosa, C. tropicalis, C. glabrata and C. utilis was strongly inhibited by aqueous extracts from the mushrooms Lentinula edodes (logs and compound), with MICs ranged varied from 0.08 to 0.31 mg / mL. These results demonstrate that edible mushrooms have great potential use by the pharmaceutical industry and can contribute significantly for a good health condition due to the presence of antioxidants
Mestrado
Ciência de Alimentos
Mestre em Ciência de Alimentos

The thesis deals with basic principles of metamaterials, which exhibit unusual properties in microwave applications (e.g., negative permittivity and permeability). Different type of metamaterial antennas and parameters of such antennas are described in the thesis.

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

碩士
國立中興大學
園藝學系
94
Petunia, Impatiens, cosmos and other annual flowers have been cultivated in the campus of National Chung Hsing University from October to next Spring. In order to protect the turfgrass of campus, spent mushroom compost (SMC) had been used as cover medium to soil of petunia and other flowers by non-tillage. Due to the higher electrical conductivity (E.C.) of SMC, the seedlings of plug can’t been planted in the SMC directly except the pottings with 3 inches. The results showed that SMC has great potential to grow annual plants. In order to protect the environment, the SMC and other abandoned mushroom compost in local area should be recycled as cultivated medium for sustainable horticulture.

by Ching Mei Lun.
Thesis (M.Phil.)--Chinese University of Hong Kong, 1997.
Includes bibliographical references (leaves 137-145).
List of Tables --- p.I
List of Figures --- p.III
Abbreviations --- p.VII
Chapter Chapter 1 --- Introduction --- p.1
Chapter 1.1 --- Present situation of municipal solid wastes in Hong Kong --- p.1
Chapter 1.2 --- Landfill in Hong Kong --- p.1
Chapter 1.2.1 --- Landfill leachate --- p.9
Chapter 1.2.1.1 --- Generation --- p.9
Chapter 1.2.1.2 --- Quality --- p.10
Chapter 1.2.1.3 --- Environmental hazard --- p.17
Chapter 1.2.1.4 --- Treatment --- p.18
Chapter 1.2.1.5 --- Other alternatives --- p.24
Chapter 1.3 --- Spent mushroom compost --- p.27
Chapter 1.3.1 --- Production and nature --- p.27
Chapter 1.3.2 --- Availability --- p.29
Chapter 1.3.3 --- Physical and chemical properties --- p.31
Chapter 1.3.4 --- Capabilities to degrade phenolic compounds --- p.31
Chapter 1.3.5 --- Potential uses --- p.37
Chapter 1.4 --- Proposal and exp erimental plan --- p.38
Chapter Chapter 2 --- Materials and Methods --- p.41
Chapter 2.1 --- Materials --- p.41
Chapter 2.2 --- Physical and chemical analyses of pollutants --- p.41
Chapter 2.3 --- Basic studies on removal capacities on potential pollutants --- p.48
Chapter 2.3.1 --- "Removal of dyes, metals and ammonia" --- p.48
Chapter 2.3.2 --- Removal of pentachlorophenol --- p.53
Chapter 2.4 --- Applied studies on removal of pollutants --- p.58
Chapter 2.4.1 --- Treatment of landfill leachate --- p.58
Chapter 2.4.2 --- Microcosm to examine the decomposition of refuse --- p.60
Chapter 2.4.3 --- Phytotoxicity --- p.65
Chapter 2.5 --- Statistical analysis --- p.65
Chapter Chapter 3 --- Results --- p.67
Chapter 3.1 --- Characterization of spent mushroom compost and landfill leachate --- p.67
Chapter 3.2 --- Removal capacities of spent mushroom compost --- p.67
Chapter 3.2.1 --- Biosorption of dyes --- p.67
Chapter 3.2.1.1 --- Evercion yellow --- p.67
Chapter 3.2.1.2 --- Evercion navy H-ER blue --- p.73
Chapter 3.2.1.3 --- Congo red --- p.74
Chapter 3.2.1.4 --- Adsorption isotherm --- p.75
Chapter 3.2.2 --- Biosorption of metals --- p.75
Chapter 3.2.2.1 --- Lead --- p.75
Chapter 3.2.2.2 --- Iron --- p.81
Chapter 3.2.2.3 --- Cadmium --- p.82
Chapter 3.2.2.4 --- Adsorption isotherm --- p.82
Chapter 3.2.3 --- Removal of ammonia --- p.85
Chapter 3.2.3.1 --- Basic study --- p.85
Chapter 3.2.3.2 --- Applied removal of ammonia from landfill leachate --- p.85
Chapter 3.2.3.2.1 --- Effect of indigenous micro-organims in landfill leachate --- p.85
Chapter 3.2.3.2.2 --- Effect of spent mushroom compost and glucose --- p.85
Chapter 3.2.3.2.3 --- Effect of sugar cane waste extract --- p.89
Chapter 3.2.3.2.4 --- Effect of sugar cane waste and concentration of glucose --- p.89
Chapter 3.2.4 --- Removal of pentachlorophenol --- p.91
Chapter 3.2.4.1 --- Removal by spent mushroom compost --- p.91
Chapter 3.2.4.2 --- Identification of two spent mushroom compost micro-organisms --- p.91
Chapter 3.2.4.3 --- Pentachlorophenol-degrading abilities of the two micro-organisms --- p.99
Chapter 3.2.5 --- A microcosm to examine the decomposition of refuse --- p.99
Chapter 3.2.5.1 --- pH --- p.99
Chapter 3.2.5.2 --- Salinity --- p.99
Chapter 3.2.5.3 --- Turbidity --- p.103
Chapter 3.2.5.4 --- Ammonia content --- p.103
Chapter 3.2.5.5 --- Orthophosphate content --- p.106
Chapter 3.2.5.6 --- "Inorganic, organic and total carbon contents" --- p.106
Chapter 3.2.5.7 --- Metals --- p.106
Chapter 3.2.5.8 --- Gases production --- p.112
Chapter 3.2.6 --- Phytotoxicity --- p.112
Chapter Chapter 4 --- Discussion --- p.117
Chapter 4.1 --- Characterization of the spent mushroom compost --- p.117
Chapter 4.2 --- Removal abilities of pollutants by the spent mushroom compost --- p.119
Chapter 4.2.1 --- Metals and dyes --- p.119
Chapter 4.2.1.1 --- Adsorption --- p.119
Chapter 4.2.1.2 --- Adsorption specificity --- p.123
Chapter 4.2.1.3 --- Adsorption isotherm --- p.125
Chapter 4.2.2 --- Pentachlorophenol --- p.127
Chapter 4.3 --- Decomposition of refuse --- p.129
Chapter 4.4 --- Removal of ammonia in landfill leachate --- p.132
Chapter 4.5 --- Phytotoxicity --- p.133
Chapter Chapter 5 --- Conclusion --- p.135
Chapter Chapter 6 --- Reference --- p.137
Chapter Chapter 7 --- Appendix --- p.146

Lau Kan Lung.
Thesis submitted in: November 2001.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2002.
Includes bibliographical references (leaves 286-312).
Abstracts in English and Chinese.
List of Symbols and Abbreviations --- p.I
List of Figures --- p.III
List of Tables --- p.XII
Chapter 1 --- Introduction --- p.1
Chapter 1.1 --- Polycyclic aromatic hydrocarbons (PAHs) --- p.1
Chapter 1.1.1 --- Physical and chemical properties of PAHs --- p.1
Chapter 1.1.2 --- Formation of PAHs --- p.5
Chapter 1.1.3 --- Sources of PAHs --- p.9
Chapter 1.1.4 --- Regulations for contamination of PAHs --- p.13
Chapter 1.1.5 --- Pollution of PAHs in environments of Hong Kong --- p.17
Chapter 1.1.6 --- Toxicity of PAHs --- p.18
Chapter 1.1.7 --- Fate of PAHs --- p.22
Chapter 1.1.7.1 --- Sorption --- p.24
Chapter 1.1.7.2 --- Volatilization --- p.25
Chapter 1.1.7.3 --- Photooxidation --- p.25
Chapter 1.1.7.4 --- Chemical oxidation --- p.27
Chapter 1.1.7.5 --- Microbial degradation --- p.28
Chapter 1.1.8 --- General principles of metabolism of PAHs --- p.30
Chapter 1.2 --- Spent mushroom compost (SMC) --- p.35
Chapter 1.2.1 --- Production of SMC --- p.35
Chapter 1.2.2 --- Physical and chemical properties of SMC --- p.36
Chapter 1.2.3 --- Availability of SMC --- p.40
Chapter 1.2.4 --- Conventional applications of SMC --- p.43
Chapter 1.2.5 --- Alternate use of SMC --- p.44
Chapter 1.3 --- Objectives of the study --- p.48
Chapter 1.4 --- Research strategy --- p.51
Chapter 1.4.1 --- Effect of initial PAH concentration --- p.51
Chapter 1.4.2 --- Effect of initial pH --- p.52
Chapter 1.4.3 --- Effect of incubation time --- p.53
Chapter 1.4.4 --- Effect of incubation temperature --- p.54
Chapter 1.4.5 --- Putative identification of intermediates and/or breakdown products --- p.54
Chapter 1.4.6 --- Isotherm plots and fitting into monolayer models --- p.55
Chapter 1.4.6.1 --- Langmuir isotherm --- p.56
Chapter 1.4.6.2 --- Freundlich isotherm --- p.58
Chapter 1.4.7 --- Toxicological study by Microtox test --- p.59
Chapter 1.4.8 --- Removal of PAH mixtures --- p.60
Chapter 1.4.9 --- Specific goals of the study --- p.61
Chapter 2 --- Materials and Methods --- p.62
Chapter 2.1 --- Materials --- p.62
Chapter 2.2 --- Physical and chemical analysis of SMC --- p.62
Chapter 2.2.1 --- pH --- p.63
Chapter 2.2.2 --- Electrical conductivity --- p.63
Chapter 2.2.3 --- Salinity --- p.63
Chapter 2.2.4 --- Ash content --- p.63
Chapter 2.2.5 --- Metal contents --- p.64
Chapter 2.2.6 --- Water-soluble anion contents --- p.65
Chapter 2.2.7 --- "Carbon, hydrogen, nitrogen and sulfur contents" --- p.65
Chapter 2.2.8 --- Infrared spectroscopic study --- p.66
Chapter 2.2.9 --- Chitin content --- p.66
Chapter 2.3 --- Soil collection and characterization --- p.67
Chapter 2.4 --- Optimization for extraction --- p.67
Chapter 2.5 --- Removal of PAHs --- p.68
Chapter 2.5.1 --- Experimental design --- p.68
Chapter 2.5.1.1 --- Pretreatment and incubation --- p.68
Chapter 2.5.1.2 --- Extraction of sorbed PAHs in soil system or in SMC --- p.69
Chapter 2.5.1.3 --- Extraction of PAHs in water system --- p.70
Chapter 2.5.1.4 --- Putative identification and quantification of PAHs --- p.71
Chapter 2.5.2 --- Assessment criteria --- p.72
Chapter 2.5.3 --- Stability of PAHs --- p.77
Chapter 2.5.4 --- Optimization for removal of PAHs --- p.78
Chapter 2.5.4.1 --- Effects of initial PAH concentration and amount of SMC --- p.78
Chapter 2.5.4.2 --- Effect of initial pH --- p.79
Chapter 2.5.4.3 --- Effect of incubation time --- p.79
Chapter 2.5.4.4 --- Effect of incubation temperature --- p.79
Chapter 2.5.5 --- Putative identification of intermediates and/or breakdown products --- p.80
Chapter 2.5.6 --- Isotherm plots and fitting into monolayer models --- p.80
Chapter 2.5.6.1 --- Langmuir isotherm --- p.80
Chapter 2.5.6.2 --- Freundlich isotherm --- p.81
Chapter 2.5.7 --- Toxicological study of Microtox® test --- p.82
Chapter 2.5.8 --- Removal ability of SMC towards PAHs in single and in a mixture --- p.82
Chapter 2.5.9 --- Removal abilities of different sorbents towards PAHs in water --- p.83
Chapter 2.5.10 --- Removal abilities of raw and autoclaved SMC towards PAHs in water --- p.83
Chapter 2.5.11 --- Statistical validation --- p.83
Chapter 3 --- Results --- p.85
Chapter 3.1 --- Characterization of soil --- p.85
Chapter 3.1.1 --- Physical and chemical properties of soil --- p.85
Chapter 3.1.2 --- GC-MS analysis of soil --- p.85
Chapter 3.2 --- Calibration curves of PAHs --- p.85
Chapter 3.3 --- Optimization for extraction --- p.91
Chapter 3.4 --- Stability of PAHs --- p.101
Chapter 3.4.1 --- Soil system --- p.101
Chapter 3.4.1.1 --- Effect of incubation time --- p.101
Chapter 3.4.1.2 --- Effect of incubation temperature --- p.101
Chapter 3.4.2 --- Water system --- p.103
Chapter 3.4.2.1 --- Effect of incubation time --- p.103
Chapter 3.4.2.2 --- Effect of incubation temperature --- p.103
Chapter 3.5 --- Characterization of SMC --- p.103
Chapter 3.5.1 --- Physical and chemical properties of SMC --- p.103
Chapter 3.5.2 --- GC-MS analysis of SMC --- p.106
Chapter 3.5.3 --- Infrared spectroscopic study and chitin content --- p.106
Chapter 3.5.4 --- Removal abilities of different sorbents towards PAHs in water --- p.121
Chapter 3.5.5 --- Removal abilities of raw and autoclaved SMC towards PAHs in water --- p.121
Chapter 3.6 --- Optimization for removal of PAHs --- p.124
Chapter 3.6.1 --- Naphthalene --- p.124
Chapter 3.6.1.1 --- Soil system --- p.124
Chapter 3.6.1.1.1 --- Effects of initial naphthalene concentration and amount of straw SMC on removal efficiency --- p.124
Chapter 3.6.1.1.2 --- Effects of initial naphthalene concentration and amount of straw SMC on removal capacity --- p.128
Chapter 3.6.1.1.3 --- Effect of initial pH --- p.128
Chapter 3.6.1.1.4 --- Effect of incubation time --- p.128
Chapter 3.6.1.1.5 --- Effect of incubation temperature --- p.131
Chapter 3.6.1.1.6 --- Putative identification of intermediates and/or breakdown products --- p.131
Chapter 3.6.1.2 --- Water system --- p.134
Chapter 3.6.1.2.1 --- Effects of initial naphthalene concentration and amount of straw SMC on removal efficiency --- p.134
Chapter 3.6.1.2.2 --- Effects of initial naphthalene concentration and amount of straw SMC on removal capacity --- p.137
Chapter 3.6.1.2.3 --- Effect of initial pH --- p.137
Chapter 3.6.1.2.4 --- Effect of incubation time --- p.139
Chapter 3.6.1.2.5 --- Effect of incubation temperature --- p.139
Chapter 3.6.1.2.6 --- Putative identification of intermediates and/or breakdown products --- p.143
Chapter 3.6.2 --- Phenanthrene --- p.145
Chapter 3.6.2.1 --- Soil system --- p.145
Chapter 3.6.2.1.1 --- Effects of initial phenanthrene concentration and amount of straw SMC on removal efficiency --- p.145
Chapter 3.6.2.1.2 --- Effects of initial phenanthrene concentration and amount of straw SMC on removal capacity --- p.145
Chapter 3.6.2.1.3 --- Effect of initial pH --- p.148
Chapter 3.6.2.1.4 --- Effect of incubation time --- p.148
Chapter 3.6.2.1.5 --- Effect of incubation temperature --- p.151
Chapter 3.6.2.1.6 --- Putative identification of intermediates and/or breakdown products --- p.151
Chapter 3.6.2.2 --- Water system --- p.155
Chapter 3.6.2.2.1 --- Effects of initial phenanthrene concentration and amount of straw SMC on removal efficiency --- p.155
Chapter 3.6.2.2.2 --- Effects of initial phenanthrene concentration and amount of straw SMC on removal capacity --- p.155
Chapter 3.6.2.2.3 --- Effect of initial pH --- p.157
Chapter 3.6.2.2.4 --- Effect of incubation time --- p.157
Chapter 3.6.2.2.5 --- Effect of incubation temperature --- p.161
Chapter 3.6.2.2.6 --- Putative identification of intermediates and/or breakdown products --- p.163
Chapter 3.6.3 --- Benzo[a]pyrene --- p.163
Chapter 3.6.3.1 --- Soil system --- p.163
Chapter 3.6.3.1.1 --- Effects of initial benzo[a]pyrene concentration and amount of straw SMC on removal efficiency --- p.163
Chapter 3.6.3.1.2 --- Effects of initial benzo[a]pyrene concentration and amount of straw SMC on removal capacity --- p.167
Chapter 3.6.3.1.3 --- Effect of initial pH --- p.167
Chapter 3.6.3.1.4 --- Effect of incubation time --- p.168
Chapter 3.6.3.1.5 --- Effect of incubation temperature --- p.168
Chapter 3.6.3.1.6 --- Putative identification of intermediates and/or breakdown products --- p.172
Chapter 3.6.3.2 --- Water system --- p.172
Chapter 3.6.3.2.1 --- Effects of initial benzo[a]pyrene concentration and amount of straw SMC on removal efficiency --- p.172
Chapter 3.6.3.2.2 --- Effects of initial benzo[a]pyrene concentration and amount of straw SMC on removal capacity --- p.176
Chapter 3.6.3.2.3 --- Effect of initial pH --- p.178
Chapter 3.6.3.2.4 --- Effect of incubation time --- p.178
Chapter 3.6.3.2.5 --- Effect of incubation temperature --- p.181
Chapter 3.6.3.2.6 --- Putative identification of intermediates and/or breakdown products --- p.183
Chapter 3.6.4 --- "Benzo[g,h,i]perylene" --- p.183
Chapter 3.6.4.1 --- Soil system --- p.183
Chapter 3.6.4.1.1 --- "Effects of initial benzo[g,h,i]perylene concentration and amount of straw SMC on removal efficiency" --- p.183
Chapter 3.6.4.1.2 --- "Effects of initial benzo[g,h,i]perylene concentration and amount of straw SMC on removal capacity" --- p.187
Chapter 3.6.4.1.3 --- Effect of initial pH --- p.187
Chapter 3.6.4.1.4 --- Effect of incubation time --- p.187
Chapter 3.6.4.1.5 --- Effect of incubation temperature --- p.189
Chapter 3.6.4.1.6 --- Putative identification of intermediates and/or breakdown products --- p.189
Chapter 3.6.4.2 --- Water system --- p.192
Chapter 3.6.4.2.1 --- "Effects of initial benzo[g,h,i]perylene concentration and amount of straw SMC on removal efficiency" --- p.192
Chapter 3.6.4.2.2 --- "Effects of initial benzo[g,h,i]perylene concentration and amount of straw SMC on removal capacity" --- p.196
Chapter 3.6.4.2.3 --- Effect of initial pH --- p.198
Chapter 3.6.4.2.4 --- Effect of incubation time --- p.198
Chapter 3.6.4.2.5 --- Effect of incubation temperature --- p.198
Chapter 3.6.4.2.6 --- Putative identification of intermediates and/or breakdown products --- p.201
Chapter 3.7 --- Isotherm plots and fitting into monolayer models --- p.205
Chapter 3.7.1 --- Sorption of naphthalene --- p.205
Chapter 3.7.2 --- Sorption of phenanthrene --- p.205
Chapter 3.7.3 --- Sorption of benzo[a]pyrene --- p.208
Chapter 3.7.4 --- "Sorption of benzo[g,h,i]perylene" --- p.208
Chapter 3.8 --- Toxicological study of Microtox test --- p.208
Chapter 3.8.1 --- Soil system --- p.214
Chapter 3.8.2 --- Water system --- p.214
Chapter 3.9 --- Operable conditions of SMC for removal of PAHs --- p.214
Chapter 3.10 --- Removal ability of SMC towards PAHs in single and in a mixture --- p.214
Chapter 3.10.1 --- Soil system --- p.216
Chapter 3.10.2 --- Water system --- p.216
Chapter 4 --- Discussion --- p.221
Chapter 4.1 --- Characterization of SMC --- p.221
Chapter 4.2 --- Removal abilities of different sorbents towards PAHs in water --- p.223
Chapter 4.3 --- Removal abilities of raw and autoclaved SMC towards PAHs in water --- p.226
Chapter 4.4 --- Extraction efficiencies of PAHs --- p.227
Chapter 4.5 --- Factors affecting removal of PAHs by SMC --- p.229
Chapter 4.5.1 --- Initial PAH concentration and amount of straw SMC --- p.229
Chapter 4.5.2 --- Initial pH --- p.237
Chapter 4.5.3 --- Incubation time --- p.237
Chapter 4.5.4 --- Incubation temperature --- p.242
Chapter 4.6 --- Putative identification of intermediates and/or breakdown products --- p.247
Chapter 4.7 --- Isotherm plots and fitting into monolayer models --- p.257
Chapter 4.8 --- Toxicological study of Microtox® test --- p.258
Chapter 4.9 --- Removal ability of SMC towards PAHs in single and in a mixture --- p.261
Chapter 4.10 --- Comparison of removal efficiencies of benzo[a]pyrene by layering and mixing of straw SMC with soil --- p.265
Chapter 4.11 --- Comparison of removal efficiencies of benzo[a]pyrene in different scales of experiment setup --- p.267
Chapter 4.12 --- Effect of age of straw SMC on removal of PAHs --- p.270
Chapter 4.13 --- Removal of benzo[a]pyrene by an aqueous extract of SMC --- p.270
Chapter 4.14 --- Advantages of using SMC in removal of PAHs --- p.273
Chapter 4.15 --- Limitations of the study --- p.278
Chapter 4.16 --- Further investigation --- p.280
Chapter 5 --- Summary --- p.282
Chapter 6 --- Conclusion --- p.285
Chapter 7 --- References --- p.286

碩士
東吳大學
微生物學系
102
Sulfamethoxazole(SMX),sulfadimethoxine(SDM)and sulfamethazine (SMZ) are sulfonamides with broad spectrum in common use. Our research investigated the ability of spent mushroom compost (SMC) in the removal of sulfonamides in sludge. Via batch experiments with different conditions, optimum system was established. The results showed that the half-life (t½) of SMX treated with SMC (without extraction) was 2.4 days, which was superior to the control group (6 days). Experiments of medium-scale bioreactors were conducted with four times addition of SMX to determine the effects of enzyme addition to the degradation rates. The half-life of SMX in groups without the addition of enzyme, groups with the addition of enzyme and control groups were 1.0-1.5 days, 1.0-2.6 days and 1.5-3.2 days, respectively. The above results implied that the addition of enzyme made no significant difference in the degradation of SMX. After medium-scale bioreactors, experiments of large-scale bioreactors were performed with six times addition of sulfonamides. The half-life of SMX, SMZ and SDM in control groups were 0.9-6.0 days, 1.8-6.0 days and 0.9-4.8 days whereas in treatment groups were 0.8-2.0 days, 0.8-10.9 days and 0.9-2.1 days, respectively. The results demonstrated that the degradation rates of SDM and SMX were higher than SMZ. However, the degradation rates of SMZ in treatment groups would be enhanced as SMZ added each time. Also, once the concentrations of sulfonamides were increased from 20mg/L to 50mg/L, the half-life of SMX, SMZ and SDM would drop. Next, the processed sludge was added into soil to perform experiments of large-scale soil bioreactors. In the soil experiments with four times addition of sulfonamides, the half-life of SMX, SMZ and SDM in control groups were 0.6-5.0 days, 1.2-9.1 days and 0.7-9.4 days whereas in treatment groups were 0.8-1.9 days, 1.2-2.8 days and 0.9-1.7 days, respectively. Although the degradation rates of control groups were higher than treatment groups in the first time addition of sulfonamides, the degradation rates of treatment groups rose in the following times of addition. Meanwhile, the degradation rates of control groups were not improved significantly. In the microbial community analysis, sequences retrieved from pyrosequencing were compared with sequences in Ribosomal Database Project (RDP). The results indicated that Rummeliibacillus spp., Nocardioides spp. , Clostridium sensu stricto spp., Streptomyces spp., Methanosarcina spp. and Edaphobacter spp.were dominated in batch experiments. On the other hand, TM7_genera_incertae_sedis spp., Methylobacillus spp., Chryseobacterium spp., Castellaniella spp. , Bdellovibrio spp., Sporocytophaga spp., Chlorobium spp., Rhizomicrobium spp., Hyphomicrobium spp. and Oligotropha spp.were major populations in the experiments of large-scale bioreactors. After data analysis, these microbes were found related to the degradation of sulfonamides. Overall, the results suggested that direct addition of SMC into sludge could not only reduce the waste but strengthen the effects in sulfonamides degradation. Our research provides perspectives in coping sulfonamides polluted soils and sludge with an economical and eco-friendly way of bioremediation.

碩士
國立中興大學
園藝學系
84
The germination of Chinese cabbage were both inhibited by the sterilized or unsterilized fresh spent winter mushroom compost（SFSWMC）（FSWMC） The inhition was released if the duration of fermentation of SWMC were more than 4 weeks. The major fungus in the SWMC was Trichoderma harzianum. and Penicillium sp. The inoculation of Trichoderma harzianum. , Penicillium sp.and Bacteria into the sterilized fresh spent winter mushroom compost had the same performance with natural fermentation for the plant growth which were acceptable. Selected chemical properties of fresh spent winter mushroom compost（FSWMC） were evaluated and compared to the properties of spent winter mushroom compost which was aged（ ASWMC） aerobically for 4 weeks or more.Both contained very high levels of soluble salts if compare with sawdust or peatmoss. Concetration of NH4-N in FSWMC was high while cons. of NO3-N in ASWMC was high. Chlorsis in young leaves are easily appeared might be caused from lower available Fe or higher Mn. The growth of Cynodon dactylon and Brassica chinesis were inhibited if FSWMC（0~3 weeks aged）were used. ASWMC（> 4 weeks aged）induced vigrous growth even more than BVB4 done. The combinations of ASWMC with others ,ex. carbonized rice hull or BVB4 might improved the crop growth.

碩士
東吳大學
微生物學系
101
Tetrabromobisphenol-A (TBBPA) is one of the most widely used brominated flame retardants (BFRs) which is environmental hormone. In this study, we investigated the removal of TBBPA using the spent mushroom compost (SMC) and assess the sludge added to the soil as a soil improver. We also used pyrosequencing technology for monitoring microbial community change in TBBPA in the sludge and soil. The remaining percentages were ND, 21%, and 6.4% for the sludge-SMC, sludge-crude extracts, sludge-microcapsules (the ratio of 1:9), after 49 days incubations, respectively. The results showed that the addition of SMC, crude extract, microcapsules to sludge could enhance TBBPA degradation, with SMC yielding a higher TBBPA degradation than the other additives in sludge. The above treatments added to the soil, remaining percentage are 13%, 21%, 14% with SMC, crude extract, microcapsules, respectively after 29 days incubations, with SMC also yielding a higher TBBPA degradation than the other additives in soil. In the bioreactor experiments, the removal rates of TBBPA in sludge were first compared between reactions with and without added SMC. The removal rates of the first addition of TBBPA with SMC was 89% after 42 day, the second addition was 97.5% after 30 days, the third addition was 87% after 30 days. The results showed that SMC would change microbial community and maintained stability of TBBPA removal rate. The above treatments added to the soil in the bioreactor, the removal rates of first addition of TBBPA was 98% after 20 days, the second addition was 95.3% after 8 days, the third addition was 92.8% after 5 days. Removal efficiencies were therefore enhanced following the second and third addition of TBBPA. The pyrosequencing technology for profiling microbial communities showed that degradation of tetrabromobisphenol A to appear related bacteria. Such us Rhodanobacter spp., Steroidobacter spp., Cupriavidus spp., Brachymonas spp., in future, we will study of add pure bacterial in TBBPA degradation experiments. This study showed that added SMC could enhance TBBPA degradation in the sludge, and added the treated the sludge to soil, also increased TBBPA degradation. SMC is easily obtained and its use reduces the disposal costs of waste removal. This research offers the feasible methods for removal of TBBPA in sludge and soil for bioremediation.

碩士
東吳大學
微生物學系
106
In this study, fungi replace the traditional bacteria-based biodegradation of emerging contaminants. Five emerging contaminants of acetaminophen, sulfamethoxazole (SMX), sulfamethazine (SMZ), sulfadimethoxine (SDM) and tetrabromobisphenol A (TBBPA) were degraded with Flammulina velutipes, Hypsizygus marmoreus, Pholiota namek, Agaricus blazei, Pleurotus ferulae and Pleurotus eryngii SMC. Test the best mushroom degradation efficiency. Further bioreactor experiments were used to further verify the feasibility of using SMC to degrade emerging contaminants in WWTP wastewater. Among the SMC of five mushrooms tested by batch experiments, their ability to degrade was P. eryngii > P. ferulae > A. blazei. However, F. velutipes, H. marmoreus and P. namek showed no degradation. In addition, the efficiency of degradation of organic pollutants by P. eryngii SMC was acetaminophen > three sulfonamides > TBBPA. The most degradative SMC obtained after batch experiments screening were screened for bioreactor degradation experiments of 10 mg/l acetaminophen, three sulfonamides and TBBPA using WWTP wastewater. The results showed that P. eryngii SMC required for different contaminants varied in volume and flow rate. The flow rate of 4.17 ml/min, 50 g of P. eryngii SMC can be immediately removed 10 mg/l Acetaminophen; sulfonamides need 100 g SMC, but SMC can only be used 1 times; TBBPA requires 20.83 ml/h, 100 g SMC and is properly filtered after filtration to effectively remove TBBPA. Therefore, we can develop green products, develop and utilize SMC for resource recovery and reuse, and provide innovative biological treatment technologies for fungal remediation to remove different pollutants in the environment, to adjust the volume of SMC, the velocity of the contaminated water, the duration of action, etc. Regenerating with SMC not only achieves the goal of energy conservation and carbon reduction, but also removes many residual chemicals in our living environment.

碩士
嘉南藥理大學
環境工程與科學系
106
Biochar with porous, hydrophobic and low density properties could provide macropore for microbial growth in soils. As large development of mushroom industry, the utilization of agricultural waste resources had attracted great attention. In this study, various types of biochars made from various SMW (mushroom, oyster mushroom and coral mushroom) with various pyrolysis temperature (300, 500 and 850℃). Furthermore, several composts were prepared from various mixing ratio of pig manure, mushroom wastes and biochars in aerobic composting treatments. The results were used to evaluate effect of biochar addition on humification and quality of compost and their application for plant growth. The biochars pyrolysed in 800℃ presented higher pH, electric conductivity, C/N ratio and BET areas. During the composting reaction, the pH and organic matter contents of composts with various biochar amendment presented no obvious differences. Composts with higher biochar addition (10%) retained low EC vaules. During the composting process, small molecules were synthesized to form stable macromolecules during composting. E4/E6 values with biochar addition were lower than non-additive case during composting. In the late stage of composting, organic compostions of materials formed to humus substances gradually and the alphatic groups disappeared by FTIR spectrum. It was indicated that biochar addition enhanced the humification and quality of compost. In the seed germination, water extractable (compost:water, 1:5) solutions of all compost products showed good germination rate. Comparison of effects of biochar composts on plant growth was taken in pot experiment of Brassica chinensis Linn with amendment of chemical ferlizer as control. Higher pH and OM contents were observed in the amendment of biochar compost than control. The best performance of plant growth which amended with 10% M500 biochar compost was observed. It was indicated that proper reuse of agricultural waste could improve soil environment and had a valuable application potential for plants harvest.

碩士
國立中興大學
植物病理學系
87
Of the four agriculture wastes tested, spent forest mushroom compost (SFMC) was more suitable as substrates for growth of cucumber seedlings than raw growth medium for cultivation of forest mushroom (RGM), raw spent forest mushroom growth medium (RFM), and raw spent forest mushroom growth medium contaminated with other microorganisms [ RFM (C) ]. Inoculation method of zoospore suspension provided a stable and accurate means of assessing the effect of SFMC and Bas Van Burren No.4 peat moss (BVB) on damping-off incidence of cucumber seedlings. SFMC was more effective in suppressing damping-off of cucumber seedlings caused by Pythium aphanidermatum isolate Pa104 and Pa105 as compared with BVB. Five SFMC-BVB medium combinations were prepared by mixing SFMC with BVB at rates of 4:0, 3:1, 2:2, 1:3, and 0:4 (v/v) and inoculated with zoospore suspension of the pathogen. The more concentration of SFMC the medium was amended, the less incidence of damping-off the cucumber seedlings were grown in the medium for 12days. SFMC was not suitable for survival of P. aphanidermatum isolates Pa104 and Pa105 because the fungi could not be detected by Corn meal-pimaricin-rose bengal-streptomycin-benomyl agar 28 days after their perlite inoculum were added into SFMC medium. The suppressive effect of SFMC medium on infection of cucumber seedlings by P. aphanidermatum isolates Pa104 & Pa105 was not reduced after it was steamed at 100℃ hot air for 20 min. or autoclaved at 121℃, 15lb, for 20 min. The water-soluble filtrate extracted from SFMC could markedly inhibit formation of oOgonia and antheridia of P. aphanidermatum isolates Pa104 & Pa105 and reduce production of zoospores as compared with BVB's water-soluble filtrate. However, filtrates from SFMC and BVB were not significantly different in affecting the formation of zoosporangia. The electrical conductivity values of SFMC filtrate were highly correlated with production of zoospores by the pathogen. The inhibitory effect of filtrate from SFMC medium were partially nullified after it was steamed and autoclaved. However, the steamed and autoclaved SFMC medium still showed adverse effect on survival of the pathogen. Cucumber seedlings grown in SFMC medium were more effective in inhibiting colonization of root, stem, and cotylendon by the pathogen than ones in BVB medium. The peroxidase activity, nutrient elements such as K, Ca, Mg, NO3--N and NH4+-N in root and stem from cucumber seedlings grown in SFMC medium were significantly higher than those in BVB medium. The results suggest that abiotic factors may play an important role on reduction of damping-off incidence of cucumber seedlings in SFMC medium.

碩士
國立中興大學
水土保持學系所
101
The application of hydroseeding allows fast planting and greening for rapidly covering landslides and reducing runoff to decrease second damage and provide basement conditions for plant growth. The application and improvement of mixed materials are continuously developed. Total 13 test recipes are selected for this study, containing four types of Portland cement (with mixed material dry weight ratio of 1％, 3％, 5％, and 7％), three types of loamy sandy soil (with mixed material volume ratio 10％, 20％, and 40％), three types of mushroom compost (with mixed material volume ratio 90％, 80％, and 60％), and the control group of 100％ mushroom compost. The mixed material are added the seeds of Lolium multiflorum and Acacia confuse for laboratory tests to discuss the effects of mixed materials with distinct ratios on the seed germination and to provide the proof and reference of mixed material application benefits. The results are summarized as below. 1.According to the physicochemical property analysis of various mixed materials recipes, the higher cement ratio would increase the pH value, hardness, and lognormal geometric mean diameter of mixed materials and enhance the erosion resistance. 2.Water retention and electrical conductivity (EC) are affected by the mushroom compost ratio that the higher ratio of mushroom compost, the better water retention and the higher electrical conductivity. The tests of electrical conductivity present that the highest EC among various recipes appears in 1.33 dS/m, which is suitable for seed germination. 3.From the test results, the germination percentage of Lolium multiflorum with various recipes reaches up to 85％, revealing that the seed germination is not significantly affected by cement ratio. In the comparison of aggregate stability analysis with wet-sieving and water retention, the erosion resistance of the recipe of mushroom compost 90％+ loamy sandy soil 10％+cement 7％ appears the highest water content after adding water for 28 days that it is the optimal recipe. The germination percentage also achieves more than 90％ that it is regarded as the best mixed material in this study.

An industrial soil collected from a recycling factory was contaminated by DEHP and spilled diesel. The soils were divided into two batches for off-site ex situ bioremediation by SMC. This industrial soil was contaminated with 5.4-6.9 g/kg total petroleum hydrocarbons (TPH), 14.5--19.0 g/kg oil and grease and 95-99 mg/kg DEHP. The removal by 3% SMC amendment applied twice accounted for 56-64%, 31--33% and 51--54% disappearance of the TPH, oil and grease and DEHP contaminants, respectively. Beside chemical analysis, six bacteria and six fungi were inoculated into the sterilized soil samples for ecotoxicity tests. The original soil samples containing residual oil and DEHP contents were found to be more toxic than the SMC-treated soil. Thus SMC simultaneously degrades organic pollutants and reduces toxicity in less than a month.
An underground water sample contaminated with dibutyl phthalate (DBP, 127.5+/-20.7 mu1/1) and DEHP (67.0+/-7.7 mu1/1) was tested. One percent SMC could remove 94.2+/-3.6% and 100% for DBP and DEHP, respectively, within 1 h at room temperature. DBP and DEHP were degraded completely by SMCE except 0.2% SMCE for DBP after 24-h treatment. SMC had higher removal efficiencies than SMCE, because SMC had an integrated system of biosorption and biodegradation to remove DBP and DEHP. The sorption kinetics of DBP and DEHP by SMC could be described by the Freundlich monolayer model. Even after eight cycles of the sorption-desorption of DEHP SMC maintained the 100% removal efficiency. Thus SMC is a good biosorbent for DEHP.
Besides, DEHP-contaminated sediment was collected from Kai Tak Approach Channel, Kowloon. This sediment contained 44.4-128.0 mg/kg DEHP and heavy metals 21.3-23.4 mg/kg Cd, 24.9-43.5 mg/kg Ni, 128.5-198.5 mg/kg Pb, 144.6-329.2 mg/kg Zn and 164.5-230.0 mg/kg Cu, and bore an unpleasant sewer smell. SMC and SMCE as strong oxidizing agents could decrease the S content of sediment significantly as well as calcium nitrate, and consequently lowered the evolution of nuisance gas hydrogen sulfide. With the treatment of the optimized combination of 2.25% SMCE and 0.25% nitrate, the malodor could be removed completely and the contents of H2S and NH3 in the air were decreased significantly. For the degradation of organic pollutants, the target pollutant DEHP showed a sharp decrease in the first week in the time effect experiment after the treatment with the combination of 2.25% SMCE and 0.25% nitrate, while the decrease of DEHP slowed down in the second week. The results may be attributed to the immediate degradation of DEHP by enzymes in SMCE. When raw sediment and nitrate-treated sediment were bioremediated with SMCE, larger fluctuation in DEHP removal was observed with the nitrate-treated sediment. It supports that nitrate and SMCE would act in optimum at a certain combination. The effect on mobilization of sediment heavy metals by SMC or SMCE was also examined. Although some laboratory results suggested reduction of copper and lead, other results using different environmental samples of the sediment did not reproduce the results. Further investigation is needed.
Plasticizers are additives used in the manufacture of plastics, and high residual plasticizer levels are encountered in the environment. Bis(2-ethylhexyl) phthalate (DEHP), being the most common plasticizer, is a suspected human carcinogen and an endocrine disruptor. Therefore, DEHP-contaminated soil, sediment and water samples were collected, and the bioremediation capacities of the spent compost of mushroom Pleurotus pulmonarius (SMC) were determined.
This study reveals the potential in applying SMC of P. pulmonarius in bioremediation of DEHP from the soil, sediment and water environments. More investigation and field studies would be appropriate for developing spent P. pulmonarius compost in environmental cleanup.
Using artificially spiked garden soil with DEHP and ultra-low sulphur diesel, the removal efficiencies of SMC were 41.7+/-9.8% and 36.1+/-8.4% for TP11 and DEHP, respectively. SMCE which contained the water-soluble nutrients, SMC enzymes and micro-organisms had about half removal efficiency of SMC on the same pollutant. The enriched SMC microorganisms also showed biodegradation of DEHP and diesel, so did the filtrate containing enzymes and nutrients of SMC. Thus SMC acted in multiple ways in bioremediation of DEHP: biostimulation and bioaugmentation. Besides, the immobilized lignolytic enzymes of the mushroom P. pulmonarius played a major role in biodegradation.
Gao, Ting.
"October 2009."
Adviser: S.W. Chiu.
Source: Dissertation Abstracts International, Volume: 72-01, Section: B, page: .
Thesis (Ph.D.)--Chinese University of Hong Kong, 2010.
Includes bibliographical references (leaves 204-237).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.

by Wai Lok Man.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2003.
Includes bibliographical references (leaves 142-155).
Abstracts in English and Chinese.
Acknowledgements --- p.i
Abstracts --- p.ii
Contents --- p.vii
List of figures --- p.xiii
List of tables --- p.xvi
Abbreviations --- p.xviii
Chapter 1. --- Introduction --- p.1
Chapter 1.1 --- Pentachlorophenol
Chapter 1.1.1 --- Applications of pentachlorophenol --- p.1
Chapter 1.1.2 --- Characteristics --- p.3
Chapter 1.1.3 --- Pentachlorophenol in the environment --- p.3
Chapter 1.1.4 --- Toxicity of Pentachlorophenol --- p.6
Chapter 1.2 --- Treatments of Pentachlorophenol --- p.10
Chapter 1.2.1 --- Physical treatment --- p.10
Chapter 1.2.2 --- Chemical treatment --- p.11
Chapter 1.2.3 --- Biological treatment --- p.13
Chapter 1.3 --- Biodegradation --- p.14
Chapter 1.3.1 --- Biodegradation of PCP by bacteria --- p.14
Chapter 1.3.2 --- Biodegradation of PCP by white-rot fungi --- p.15
Chapter 1.4 --- Biosorption --- p.24
Chapter 1.5 --- Proposed Strategy --- p.28
Chapter 1.6 --- Spent Mushroom Compost
Chapter 1.6.1 --- Background --- p.28
Chapter 1.6.2 --- Physico-chemical properties of SMC --- p.29
Chapter 1.6.3 --- As a biosorbent --- p.29
Chapter 1.6.3.1 --- Factors affecting biosorption --- p.31
Chapter 1.6.3.2 --- Contact time --- p.31
Chapter 1.6.3.3 --- Initial pH --- p.32
Chapter 1.6.3.4 --- Concentration of biosorbent --- p.33
Chapter 1.6.3.5 --- Initial PCP concentration --- p.34
Chapter 1.6.3.6 --- Incubation temperature --- p.34
Chapter 1.6.3.7 --- Agitation speed --- p.35
Chapter 1.6.4 --- Modeling of adsorption --- p.36
Chapter 1.6.4.1 --- Langmuir isotherm --- p.36
Chapter 1.6.4.2 --- Freundlich isotherm --- p.36
Chapter 1.6.5 --- As a source of PCP-degrading bacteria --- p.38
Chapter 1.6.5.1 --- Identification of PCP-degrading bacterium --- p.40
Chapter 1.6.6 --- As a source of fungus --- p.42
Chapter 1.7 --- Objectives of this Study --- p.43
Chapter 2. --- Materials and Methods --- p.44
Chapter 2.1 --- Spent Mushroom compost (SMC) Production --- p.44
Chapter 2.2 --- Characterization of SMC --- p.46
Chapter 2.2.1 --- pH --- p.46
Chapter 2.2.2 --- Electrical conductivity --- p.46
Chapter 2.2.3 --- "Carbon, hydrogen, nitrogen and sulphur contents" --- p.46
Chapter 2.2.4 --- Infrared spectroscopic study --- p.47
Chapter 2.2.5 --- Metal analysis --- p.47
Chapter 2.2.6 --- Anion content --- p.47
Chapter 2.2.7. --- Chitin assay --- p.48
Chapter 2.3 --- Extraction of PCP --- p.49
Chapter 2.3.1 --- Selection of extraction solvent --- p.49
Chapter 2.3.2 --- Selection of desorbing agent --- p.49
Chapter 2.3.3 --- Extraction efficiency --- p.50
Chapter 2.4 --- Adsorption of Pentachlorophenol on SMC --- p.50
Chapter 2.4.1 --- Preparation of pentachlorophenol (PCP) stock solution --- p.50
Chapter 2.4.2 --- Batch adsorption experiment --- p.51
Chapter 2.4.3 --- Quantification of PCP by HPLC --- p.51
Chapter 2.4.4 --- Data analysis for biosorption --- p.51
Chapter 2.4.5 --- Optimization of PCP adsorption --- p.52
Chapter 2.4.5.1 --- Effect of contact time --- p.52
Chapter 2.4.5.2 --- Effect of initial pH --- p.52
Chapter 2.4.5.3 --- Effect of incubation temperature --- p.53
Chapter 2.4.5.4 --- Effect of shaking speed --- p.53
Chapter 2.4.5.5 --- Effect of initial PCP concentration and amount of biosorbent --- p.53
Chapter 2.4.6 --- Adsorption isotherm --- p.53
Chapter 2.4.7 --- Effect of removal efficiency on reuse of biosorbent --- p.54
Chapter 2.5 --- Biodegradation by Isolated Bacterium --- p.54
Chapter 2.5.1 --- Isolation of PCP-tolerant bacteria from mushroom compost --- p.54
Chapter 2.5.2 --- Screening for the best PCP-tolerant bacterium --- p.54
Chapter 2.5.3 --- Identification of the isolated bacterium --- p.55
Chapter 2.5.3.1 --- 16S ribosomal DNA sequencing --- p.55
Chapter 2.5.3.1.1 --- Extraction of DNA --- p.55
Chapter 2.5.3.1.2 --- Specific PCR for 16S rDNA --- p.56
Chapter 2.5.3.1.3 --- Gel electrophoresis --- p.57
Chapter 2.5.3.1.4 --- Purification of PCR products --- p.57
Chapter 2.5.3.1.5 --- Sequencing of 16S rDNA --- p.58
Chapter 2.5.3.2 --- Gram staining --- p.60
Chapter 2.5.3.3 --- Biolog Microstation System --- p.60
Chapter 2.5.3.4 --- MIDI Sherlock Microbial Identification System --- p.61
Chapter 2.5.4 --- Optimization of PCP degradation by PCP-degrading bacterium --- p.62
Chapter 2.5.4.1 --- Effect of incubation time --- p.63
Chapter 2.5.4.2 --- Effect of shaking speed --- p.63
Chapter 2.5.4.3 --- Effect of initial PCP concentration and inoculum size --- p.63
Chapter 2.5.4.4 --- Study of PCP degradation pathway by isolated bacterium using GC-MS --- p.64
Chapter 2.6 --- Biodegradation by Fungus Pleurotus pulmonarius --- p.64
Chapter 2.6.1 --- Optimization of PCP degradation by P. pulmonarius --- p.65
Chapter 2.6.1.1 --- Effect of incubation time --- p.65
Chapter 2.6.1.2 --- Effect of shaking speed --- p.65
Chapter 2.6.1.3 --- Effect of initial PCP concentration and inoculum size --- p.65
Chapter 2.6.2 --- Study of PCP degradation pathway by fungus using GC-MS --- p.65
Chapter 2.6.3 --- Specific enzyme assays --- p.66
Chapter 2.6.3.1 --- Extraction of protein and enzymes --- p.66
Chapter 2.6.3.2 --- Protein --- p.66
Chapter 2.6.3.3 --- Laccase --- p.67
Chapter 2.6.3.4 --- Manganese peroxidase (MnP) --- p.67
Chapter 2.6.4 --- Microtox® assay --- p.67
Chapter 2.7 --- Statistical Analysis --- p.68
Chapter 3. --- Results --- p.69
Chapter 3.1 --- Physico-chemical Properties of SMC --- p.69
Chapter 3.2 --- Extraction Efficiency and Desorption Efficiency of PCP --- p.69
Chapter 3.3 --- Batch Adsorption Experiments --- p.76
Chapter 3.3.1 --- Optimization of adsorption conditions --- p.76
Chapter 3.3.1.1 --- Effect of contact time --- p.76
Chapter 3.3.1.2 --- Effect of initial pH --- p.76
Chapter 3.3.1.3 --- Effect of shaking speed --- p.79
Chapter 3.3.1.4 --- Effect of incubation temperature --- p.79
Chapter 3.3.1.5 --- Effect of initial PCP concentration and amount of biosorbent --- p.79
Chapter 3.3.2 --- Reuse of SMC --- p.83
Chapter 3.3.3 --- Isotherm plot --- p.83
Chapter 3.4 --- Biodegradation by PCP-degrading Bacterium --- p.86
Chapter 3.4.1 --- Isolation and purification of PCP-tolerant bacteria --- p.86
Chapter 3.4.2 --- Identification of the isolated bacterium --- p.90
Chapter 3.4.2.1 --- 16S rDNA sequencing --- p.90
Chapter 3.4.2.2 --- Gram staining --- p.90
Chapter 3.4.2.3 --- Biolog MicroPlates Identification System --- p.90
Chapter 3.4.2.4 --- MIDI Sherlock Microbial Identification System --- p.90
Chapter 3.4.3 --- Growth curve of PCP-degrading bacterium --- p.90
Chapter 3.4.4 --- Optimization of PCP degradation by PCP-degrading bacterium --- p.97
Chapter 3.4.4.1 --- Effect of incubation time --- p.97
Chapter 3.4.4.2 --- Effect of shaking speed --- p.97
Chapter 3.4.4.3 --- Effect of initial PCP concentration and inoculum size of bacterium --- p.101
Chapter 3.4.5 --- Determination of breakdown products of PCP by PCP-degrading bacterium --- p.101
Chapter 3.5 --- Biodegradation by Fungus Pleurotus pulmonarius --- p.103
Chapter 3.5.1 --- Growth curve of P. pulmonarius --- p.103
Chapter 3.5.2 --- Optimization of PCP degradation by P. pulmonarius --- p.103
Chapter 3.5.2.1 --- Effect of incubation time --- p.103
Chapter 3.5.2.2 --- Effect of shaking speed --- p.103
Chapter 3.5.2.3 --- Effect of initial PCP concentration and inoculum size of fungus --- p.108
Chapter 3.5.3 --- Determination of breakdown products of PCP by P. pulmonarius --- p.108
Chapter 3.5.4 --- Enzyme assays --- p.108
Chapter 3.6 --- Integration of Biosorption by SMC and Biodegradation by P. pulmonarius --- p.112
Chapter 3.6.1 --- Evaluation of PCP removal by an integration system --- p.112
Chapter 3.6.2 --- Evaluation of toxicity by Micortox® assays --- p.112
Chapter 4. --- Discussion --- p.115
Chapter 4.1 --- Physico-chemical Properties of SMC --- p.115
Chapter 4.2 --- Extraction Efficiency and Desorption Efficiency of PCP --- p.116
Chapter 4.3 --- Batch Biosorption Experiment --- p.117
Chapter 4.3.1 --- Effect of contact time --- p.117
Chapter 4.3.2 --- Effect of initial pH --- p.118
Chapter 4.3.3 --- Effect of shaking speed --- p.120
Chapter 4.3.4 --- Effect of incubation temperature --- p.120
Chapter 4.3.5 --- Effect of initial PCP concentration and amount of biosorbent --- p.121
Chapter 4.3.6 --- Reuse of SMC --- p.122
Chapter 4.3.7 --- Modeling of biosorption --- p.122
Chapter 4.4 --- Biodegradation of PCP by PCP-degrading Bacterium --- p.124
Chapter 4.4.1 --- Isolation and purification of PCP-tolerant bacterium --- p.124
Chapter 4.4.2 --- Identification of the isolated bacterium --- p.125
Chapter 4.4.3 --- Optimization of PCP degradation by PCP-degrading bacterium --- p.126
Chapter 4.4.3.1 --- Effect of incubation time --- p.126
Chapter 4.4.3.2 --- Effect of shaking speed --- p.128
Chapter 4.4.3.3 --- Effect of initial PCP concentration and inoculum size of bacterium --- p.128
Chapter 4.4.4 --- PCP degradation pathway by S. marcescens --- p.129
Chapter 4.5 --- Biodegradation of PCP by Pleurotus pulmonarius --- p.130
Chapter 4.5.1 --- Optimization of PCP degradation by P. pulmonarius --- p.130
Chapter 4.5.1.1 --- Effect of incubation time --- p.131
Chapter 4.5.1.2 --- Effect of shaking speed --- p.131
Chapter 4.5.1.3 --- Effect of initial PCP concentration and inoculum size of fungus --- p.131
Chapter 4.5.2 --- Enzyme activities --- p.132
Chapter 4.5.3 --- PCP degradation pathway by P. pulmonarius --- p.133
Chapter 4.6 --- Comparison of PCP Degradation between S.marcescens and P. pulmonarius --- p.133
Chapter 4.7 --- Integration of Biosorption by SMC and Biodegradation by P. pulmonarius --- p.135
Chapter 4.8 --- Evaluation of toxicity by Microtox® assay --- p.135
Chapter 4.9 --- Comparison of PCP Removal by Integration System of Sorption and Fungal Biodegradation and Conventional Treatments --- p.136
Chapter 4.10 --- Further Investigations --- p.137
Chapter 5. --- Conclusion --- p.139
Chapter 6. --- References --- p.142

Tsang Yiu-Yuen.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2004.
Includes bibliographical references (leaves 198-218).
Abstracts in English and Chinese.
Abstract --- p.i
摘要 --- p.iii
Acknowledgments --- p.v
Table of contents --- p.vi
List of figures --- p.xi
List of tables --- p.xvi
Abbreviations --- p.xviii
Chapter 1. --- Introduction
Chapter 1.1 --- Organic pollutant and environment --- p.1
Chapter 1.2 --- Polycyclic aromatic hydrocarbon --- p.3
Chapter 1.2.1 --- Distributions and treatment standards of two target PAHs --- p.5
Chapter 1.3 --- Pentachlorophenol --- p.8
Chapter 1.3.1 --- Distribution and treatment standard of PCP --- p.10
Chapter 1.4 --- Dichlorodiphenyltrichloroethane --- p.12
Chapter 1.4.1 --- Distribution and treatment standard of DDT --- p.13
Chapter 1.5 --- Indigo carmine --- p.15
Chapter 1.6 --- Cleanup technologies towards organopollutants --- p.16
Chapter 1.6.1 --- Treatment methods for organopollutants --- p.16
Chapter 1.6.2 --- Enzyme technology on environmental cleanup --- p.18
Chapter 1.6.3 --- Oxidoreductase --- p.19
Chapter 1.6.4 --- Enzyme preparation --- p.20
Chapter 1.6.5 --- Spent mushroom compost --- p.21
Chapter 1.6.5.1 --- Laccase --- p.22
Chapter 1.6.5.2 --- Catalytic cycle of laccase --- p.23
Chapter 1.6.5.3 --- Lignin peroxidase --- p.25
Chapter 1.6.5.4 --- Catalytic cycle of LiP --- p.26
Chapter 1.6.5.5. --- Manganese peroxidase --- p.27
Chapter 1.6.5.6 --- Catalytic cycle of MnP --- p.28
Chapter 1.6.6 --- Limitations on enzyme technology --- p.29
Chapter 1.6.7 --- Enhancement of laccase activity and/or catalytic lifetime --- p.30
Chapter 1.6.8 --- Enhancement of MnP activity and/or catalytic lifetime --- p.32
Chapter 1.6.9 --- Other general approaches to maintain enzyme activity --- p.34
Chapter 1.7 --- Aims of my study --- p.35
Chapter 2. --- Materials and Methods
Chapter 2.1 --- Materials --- p.36
Chapter 2.1.1 --- Production of spent mushroom compost (SMC) --- p.36
Chapter 2.2 --- Effect of age and batches of SMCs on enzyme qualities --- p.37
Chapter 2.3 --- Maximization of enzymes extracted from SMC --- p.38
Chapter 2.3.1 --- Effect of extraction solution type --- p.38
Chapter 2.3.2 --- Effect of extraction volume --- p.39
Chapter 2.3.3 --- Effect of extraction time --- p.39
Chapter 2.3.4 --- Effect of rotation speed --- p.39
Chapter 2.4 --- Enzyme and protein quality --- p.39
Chapter 2.4.1 --- Protein assay --- p.39
Chapter 2.4.2 --- Laccase assay --- p.40
Chapter 2.4.3 --- Manganese peroxidase assay --- p.40
Chapter 2.4.4 --- Lignin peroxidase assay --- p.41
Chapter 2.4.5 --- p-glucanase assay --- p.41
Chapter 2.4.6 --- Carboxymethylcellulase assay --- p.42
Chapter 2.4.7 --- Xylanase assay --- p.42
Chapter 2.4.8 --- Lipase assay --- p.43
Chapter 2.4.9 --- Protease assay --- p.43
Chapter 2.5 --- Freeze-drying on crude enzyme preparation --- p.44
Chapter 2.5.1 --- Effect of freeze-drying --- p.44
Chapter 2.6 --- Partial purification on crude enzyme preparation --- p.44
Chapter 2.6.1 --- PAGE analyses on Pleurotus SMC's laccase and MnP --- p.44
Chapter 2.6.2 --- Effect of dialysis --- p.45
Chapter 2.7 --- Characterization of crude enzyme powder --- p.46
Chapter 2.7.1 --- Metal analysis --- p.46
Chapter 2.7.2 --- Anion contents --- p.47
Chapter 2.7.3 --- H202 content --- p.47
Chapter 2.8 --- Stability of crude enzyme at storage --- p.48
Chapter 2.9 --- Optimization of crude enzyme activities --- p.48
Chapter 2.9.1 --- Ligninolytic enzyme --- p.48
Chapter 2.9.1.1 --- Crude enzyme amount --- p.48
Chapter 2.9.1.2 --- pH effect --- p.49
Chapter 2.9.1.3 --- Temperature effect --- p.49
Chapter 2.9.1.4 --- EDTA addition --- p.49
Chapter 2.9.1.5 --- Copper ion addition --- p.49
Chapter 2.9.1.6 --- Manganese ion addition --- p.50
Chapter 2.9.1.7 --- Hydrogen peroxide addition --- p.50
Chapter 2.9.1.8 --- Malonic acid addition --- p.50
Chapter 2.9.2 --- "Other enzymes (beta-glucanase, carboxymethylcellulase and xylanase)" --- p.51
Chapter 2.9.2.1 --- Temperature effect --- p.51
Chapter 2.9.2.2 --- pH effect --- p.51
Chapter 2.10 --- Studies on the degradation ability of crude enzyme towards organopollutants --- p.51
Chapter 2.10.1 --- Removal of PAH (naphthalene and phenanthrene) --- p.52
Chapter 2.10.1.1 --- Experimental setup --- p.52
Chapter 2.10.1.2 --- Effect of PAH concentration --- p.53
Chapter 2.10.1.3 --- Effect of ABTS addition --- p.54
Chapter 2.10.1.4 --- Effect of incubation time --- p.54
Chapter 2.10.1.5 --- Putative identification and quantification of PAHs --- p.54
Chapter 2.10.2 --- Removal of pentachlorophenol --- p.56
Chapter 2.10.2.1 --- Experimental setup --- p.56
Chapter 2.10.2.2 --- Effect of PCP concentration --- p.57
Chapter 2.10.2.3 --- Effect ofABTS addition --- p.57
Chapter 2.10.2.4 --- Effect of incubation time --- p.57
Chapter 2.10.2.5 --- Putative identification and quantification of PCP --- p.57
Chapter 2.10.3 --- "Removal of 4,4´ة-DDT" --- p.58
Chapter 2.10.3.1 --- Experimental setup --- p.58
Chapter 2.10.3.2 --- Effect of DDT concentration --- p.59
Chapter 2.10.3.3 --- Effect ofABTS addition --- p.59
Chapter 2.10.3.4 --- Effect of incubation time --- p.59
Chapter 2.10.3.5 --- Putative identification and quantification of DDT --- p.60
Chapter 2.10.4 --- Removal of dye ´ؤ Indigo carmine --- p.61
Chapter 2.10.4.1 --- Experimental setup --- p.61
Chapter 2.10.4.2 --- Effect of dye concentration --- p.62
Chapter 2.10.4.3 --- Effect of ABTS addition --- p.62
Chapter 2.10.4.4 --- Effect of incubation time --- p.62
Chapter 2.11 --- Assessment criteria --- p.62
Chapter 2.11.1 --- Degradation ability --- p.62
Chapter 2.11.2 --- Toxicity of treated samples (Microtox® test) --- p.63
Chapter 2.12 --- Statistical analysis --- p.64
Chapter 3. --- Results
Chapter 3.1 --- The best SMC for enzyme preparation --- p.65
Chapter 3.2 --- Maximization of enzymes extracted from SMC --- p.72
Chapter 3.2.1 --- Effect of extraction solution type and volume on crude enzyme recovery --- p.72
Chapter 3.2.2 --- Effect of extraction time on crude enzyme recovery --- p.79
Chapter 3.2.3 --- Effect of rotation speed on crude enzyme recovery --- p.79
Chapter 3.3 --- Effect of dialysis on crude enzyme preparation --- p.82
Chapter 3.4 --- Freeze-drying on crude enzyme preparation --- p.82
Chapter 3.5 --- Characterization of crude enzyme powder --- p.86
Chapter 3.6 --- Optimization of crude enzyme activities --- p.87
Chapter 3.7 --- Storage stability of crude enzyme in powder form and liquid form --- p.115
Chapter 3.8 --- Studies on degradation ability of crude enzyme towards organopollutants --- p.135
Chapter 3.8.1 --- Degradation of naphthalene (NAP) by crude enzyme solution --- p.135
Chapter 3.8.2 --- Degradation of phenanthrene (PHE) by crude enzyme solution. --- p.141
Chapter 3.8.3 --- Degradation of pentachlorphenol (PCP) by crude enzyme solution --- p.147
Chapter 3.8.4 --- "Degradation of 4,4´ة-DDT by crude enzyme solution" --- p.152
Chapter 3.8.5 --- Degradation of Indigo carmine by crude enzyme solution --- p.158
Chapter 4. --- Discussion
Chapter 4.1 --- The best SMC for enzyme preparation --- p.163
Chapter 4.2 --- Maximization of ligninolytic enzymes extracted from SMC --- p.168
Chapter 4.2.1 --- Effect of extraction solution type and volume on crude enzyme recovery --- p.168
Chapter 4.2.2 --- Effect of extraction time on crude enzyme recovery --- p.169
Chapter 4.2.3 --- Effect of rotation speed on crude enzyme recovery --- p.169
Chapter 4.3 --- Effect of dialysis on crude enzyme extract --- p.171
Chapter 4.4 --- Freeze-drying on crude enzyme extract --- p.171
Chapter 4.5 --- Characterization of crude enzyme powder --- p.172
Chapter 4.6 --- Optimization of crude enzyme activities --- p.173
Chapter 4.6.1 --- Effect of crude enzyme amount --- p.173
Chapter 4.6.2 --- Effect of incubation pH --- p.174
Chapter 4.6.3 --- Effect of incubation temperature --- p.176
Chapter 4.6.4 --- Effect of EDTA addition --- p.177
Chapter 4.6.5 --- Effect of copper and manganese ion addition --- p.177
Chapter 4.6.6 --- Effect of hydrogen peroxide addition --- p.179
Chapter 4.6.7 --- Effect of malonic acid on maintaining enzyme activities --- p.180
Chapter 4.6.8 --- Activities and stabilities of ligninolytic enzymes under the combined optimal conditions --- p.181
Chapter 4.7 --- Storage stability of crude enzyme in powder form and liquid form --- p.182
Chapter 4.7.1 --- "β-glucanase, carboxymethylcellulase (CMCase) and xylanase activities" --- p.182
Chapter 4.7.2 --- Protein content --- p.182
Chapter 4.7.3 --- Laccase activity --- p.183
Chapter 4.7.4 --- MnP activity --- p.183
Chapter 4.8 --- Studies on the degradation ability of crude enzyme towards organopollutants --- p.185
Chapter 4.8.1 --- Degradation of naphthalene (NAP) by crude enzyme solution --- p.185
Chapter 4.8.2 --- Degradation of phenanthrene (PHE) by crude enzyme solution. --- p.187
Chapter 4.8.3 --- Degradation of pentachlorophenol (PCP) by crude enzyme solution --- p.189
Chapter 4.8.4 --- "Degradation of 4,4-DDT by crude enzyme solution" --- p.190
Chapter 4.8.5 --- Degradation of Indigo carmine by crude enzyme solution --- p.191
Chapter 4.9 --- Prospect for SMC as a source of organopollutant-degrading enzyme --- p.193
Chapter 5. --- Conclusions --- p.195
Chapter 6. --- Further Investigation --- p.197
Chapter 7. --- References --- p.198

碩士
國立中興大學
植物病理學系
85
The tap and lateral roots of garden pea seedlings (cv. Taichung 11)could be significantly injured and inhibited by butachlor [2-chloro-2',6'-diethyl-N-(butoxymethyl)-acetanilide] in the soil. Using gardenpea seedlings to bioassay the toxicity of butachlor, the discolorationpercentage of their tap roots was 13.73, 84.31 or 100 when the seedswere respectively sown in soils pre-treated with 10, 50, or 100ppm ofbutachlor for one month. Amendment of soil with 5%(w/w) spent goldenmushroom compost(SGMC) was able to significantly promote the growthand prevent roots of garden pea seedlings from injury incited bybutachlor. Amendent of Tali or Chihu soils with either Tsaohu orWufeng SGMC was effective in gradually promoting the growth ofseedlings with increasing of SGMC concentrations from 1 to 5%(w/w).The effectiveness of Tsaohu SGMC was better than Wufeng SGMC. The sameeffect occured in Tali and Chihu soils containing 50ppm butachlorafter amendment of soil with Tsaochu or Wufeng SGMC. The SGMC was ableto stimulate proliferation of microbial populations, especially Mucorspp., Penicillium spp., Trichoderma spp., Streptomyces spp., andBacillus spp. in soil with or without butachlor. Determining microbialactivity in soil containing 0-5%(w/w) SGMC or 0-1000ppm butachlor byspectrophotometric determination of the hydrolysis of fluoresceindiacetate( FDA). Hydrolysis was found to increase with increasing thedoses of butachlor, excepted for 1000ppm, and concentrations of SGMC.Amendment of infested or disinfested soil containing 50ppm butachlorwith non-sterilized SGMC could reduce 22% or more injury severity oftap roots of garden pea compared to amendment with sterilized SGMC.Amendment of soil with F-101 microbial compost, sterilized SGMCcolonized by Mucor sp.(F-101) for 7 days at 28 ℃, was the sameeffective in alleviating root injury of garden pea seedlings bybutachlor as amendment with non-sterilized SGMC. Among manypredominant microorganisms, Mucor sp.( F-101), Trichodermahamatum(F-104), T. aureoviride(F-108), T. koningii(F-115), Penicilliumverrucosum Dierckx, var. cyclopium(F-116), Streptomyces sp.(A-107),Streptomyces sp.( A-112), and Bacillus brevis(B-109) were able todegrade butachlor in amended soil, potato dextrose broth(PDB), ornutrient broth(NB). Especially, Mucor sp.(F-101), Trichodermahamatum(F-104), Trichoderma aureoviride( F-108), Trichodermakoningii(F-115) and Penicillium verrucosum var. cyclopium(F-116) werebetter than others in ability to degrade butachlor. It was found thatthe residue of butachlor markedly decreased with increment of sporeconcentration when spore suspensions(0~5 × 10^8cfu/ml) of Mucorsp.(F-101) were inoculated into sterilized soil and PDB containing50ppm butachlor and incubated for 7 days.

碩士
東吳大學
微生物學系
107
The purpose of this study is to use the spent mushroom compost(SMC) in the shiitake bags to degrade phthalates in emerging pollutants. Using the four kinds of mushrooms, Pleurotus geesteranus, Pleurotus djamor, Pleurotus eryngii, and Auricularia auricula-judae to treat kinds of phthalates, such as Benzyl butyl phthalate(BBP), Di-n-butyl phthalate(DBP), Dicyclohexyl phthalate(DCP), Diethyl phthalate(DEP) and Di-(2-ethylhexyl) phthalate(DEHP). It is testing whether the SMC can degrade phthalatesor not. Furthermore, bioreactor experiments were carried out to verify the feasibility of using the SMC to degrade the phthalate esters in the waste water from sewage treatment plant. The results for finding the best batch condition experiment results show that the SMC incubated at 25 °C with water configuration, the enzyme activity has higher enzyme activity and better degradation ability. The best condition batch experiment results show that the degradation ability in low PAEs concentration (5 mg/L) is P. eryngii > A. auricula-judae > P. geesteranus = P. djamor . In high concentration (50 mg/L), the ability to degrade PAEs is P. geesteranus > P. djamor > A. auricula-judae > P. eryngii . Four kinds of SMC could effectively remove BBP, DBP and DEP, but there was no obvious degradation or removal effect on DCP and DEHP. However, after adding the six mediators of the Laccase promoter and the six metal ions or organic acids of the lignin degrading enzyme promoter, most of them did not promote degradation. On the contrary, BBP, DBP and DEP which are degradable originally have inhibited degradation. The DCP and DEHP, which are difficult to degrade are not degraded by additives. The results show that the degradation of the phthalates may not be due to the action of Laccase. The experiment using pure Laccase was selected from three different sources, Aspergillus sp., R. vernicifera, T. versicolor, and the mediator Acetosyringone was added. However, there is no phthalates degradation with or without the addition of mediator. The above experiment can confirm that the degradation of the phthalates is not due to the action of Laccase. The bioreactor experiments were divided into ultra-pure water and waste water, and the flow rate was controlled at 16.66 and 4.17 mL/min. The results show that ultra-pure water can partially remove BBP and DBP in water, regardless of the flow rate, while DEP can only be partially removed by A. auricula-judae. BBP, DBP and DEP can be removed by the SMC, and A. auricula-judae has better outcome. In the waste water, BBP and DBP can be partially removed. Among them, the effect of the P. geesteranus is poor, and DEP has no removal effect. The SMC can completely remove BBP, DBP and DEP, and A. auricula-judae is slower. This study can be found that SMC from Pleurotus geesteranus, Pleurotus djamor, Pleurotus eryngii, and Auricularia auricula-judae can remove BBP, DBP and DEP without adding substances. In the future, the SMCs can be recovered and reused, and a new biological remediation treatment technology such as biological filter material can be provided to remove the residues of phthalates in the environment. Not only can energy saving and carbon reduction be achieved, but also environmental pollutants can be removed.

碩士
東吳大學
微生物學系
107
Emerging contaminants are the target of many studies today, and the use of biodegradation to remove contaminants has become a common method used today which is less harmful to the environment.The purpose of this study is to use the enzymes produced in the matrix after fungal cultivation to degrade UV filters that are difficult to degrade in emerging pollutants.It can not only replace the traditional biodegradation technology based on bacteria, but also achieve the effect of recycling.Among them, three kinds of SMC are used: Pleurotus eryngii, Pleurotus djamor, and Auricularia polytricha. The SMC is test for 7 kinds of UV filters (BC, OC, EHS, HMS, MBC, OBZ, EHMC) whether they can be degraded and analyze the optimal conditions for degradation.Further application of the best degradation conditions in the bioreactor experiment proves the feasibility of using the waste space package to degrade the UV filters in the sewage treatment plant wastewater. The results of the batch experiments showed that the extracts of the substrate after three SMCs were degraded for seven UV filters (BC, OC, EHS, HMS, MBC, OBZ, EHMC) and six redox mediators (ACE, SA, ABTS, HP, VA, TEMPO) have different roles, like EHS, HMS, EHMC, three kinds of SMCs can degrade them very effectively, and the overall degradation ability ranks is Pleurotus djamor >Pleurotus eryngii> Auricularia polytricha.At 2 mg/L, the degradation of UV filters with Auricularia polytricha SMCs is best.At 20 mg/L, it is better to use Pleurotus djamor. For the other four refractory UV filters (BC, OC, MBC, OBZ), it is necessary to add redox mediators to promote decomposition.It is shown that ACE can promote the removal of BC by three kinds of SMCs;ABTS can best promote the removal of OC by three kinds of SMCs;TEMPO can promote the removal of MBC by three kinds of SMCs;ABTS can best promote the removal of OBZ by three kinds of SMCs.In terms of overall degradation ability, degradation is best after adding ABTS. It is also known that after the extracts of the three kinds of mushrooms are degraded by different combinations,The pH value is higher than that before degradation;Enzyme activity is due to an increase in pH, so most of it is lower than the degradation. After comparing the extract of SMCs , the degradation experiments of redox mediators were added to 7 different concentrations of UV filter.After ten days of degradation, it can be known that in three different concentrations of UV filters, without adding ABTS, the ability of extract of Pleurotus djamor SMCs to degrade 7 kinds of UV filters is optimal.And after adding ABTS, it has better degradation ability, especially for OBZ, which has a relatively high promotion effect.Overall, Pleurotus eryngii was the best for the degradation of seven UV filters after the addition of ABTS. The results of the extracts of the three kinds of SMCs to degrade different concentrations of UV filters also showed that in 2, 10 mg / L, the extracts of the three kinds of SMCs had better degradation effect on EHS, HMS and EHMC.At 100 mg/L, it is better to degrade OC.After the addition of ABTS, the extracts of the three kinds of SMCs all have the effect of degrading OBZ at different concentrations, while the low concentration has a slight effect on other UV filters.However, at high concentrations, it did not promote the degradation of other UV filters by the extracts of the three kinds of SMCs.In this experiment, the pH value, enzyme activity, and ORP value before and after degradation were also measured. It was found that the pH value and enzyme activity were consistent with the results of the above two experiments, and the ORP value also showed that the redox potential was degraded after the degradation. After the action, it decreased, and it was found that the ORP value increased after the addition of ABTS, which also represented the action of the contained enzyme.At the same time, adsorption experiments were also carried out. The adsorption capacities of the seven UV filters were Pleurotus eryngii > Pleurotus djamor > Auricularia polytricha.Therefore, the entire batch experiment also established the optimal removal conditions for use in subsequent bioreactors. The bioreactor is provided with six combinations, which are three reactor combinations of ultrapure water and discharge water. (1) 2 mg/L 1000 mL of 7 kinds of UV filter with a water retention time of 4 hours, a capacity of 200 g of SMCs.(2) 2 mg/L 500 mL of 7 kinds of UV filters have a water retention time of 2 hours, a capacity of 100 g of SMCs.(3) 7 mg/L 500 mL of 7 kinds of UV filters have a water retention time of 30 minutes, and the capacity is 100 g of SMCs.In ultrapure water, bioreactor combination one or two, Auricularia polytricha has the best degradation ability for the seven UV filters after cultivation.The bioreactor combination three is the best degradation ability of the SMCs of Pleurotus djamor.In the bioreactor combination of the discharged water, it can be known that the degradation ability of the SMCs of the Pleurotus djamor is optimal;In the second and third bioreactor combinations, the degradation ability of the SMCs of Pleurotus eryngii was the best. The effect of adding ABTS on 7 UV filters was also tested in the Pleurotus djamor bioreactor. The results showed that the degradation of 7 UV filters by ABTS did not greatly promote the effect.It was also found that there was no significant decrease in the pH value after two days, and it was maintained at around 6, and the pH value of the added ABTS was maintained below 6. It was also found that the enzyme activity of adding 7 kinds of UV filters in ultrapure water increased after two days, but the enzyme activity of adding 7 kinds of UV filters in the discharged water decreased with or without ABTS, and the reason was unknown. In the algae biotoxicity test, it can be known that three 2 mg/L UV filters (EHS, HMS, EHMC) can reduce the survival rate of chlorella.However, after degradation by the pink mushroom extract, the toxicity is reduced and the chlorella is maintained at a certain amount.It indicates that the pink mushroom does decompose three kinds of 2 mg/L UV filter to reduce its toxicity.As a result of its metabolites, it can be known that after degradation, three kinds of 2 mg/L UV filters (EHS, HMS, EHMC) will be decomposed into other products. As for whether other products are toxic, further research is needed to do it. At the end of the study, we can know that the SMCs of Pleurotus eryngii, Pleurotus djamor and Auricularia polytricha can degrade the UV filters. This study can be used as a biodegradable method in the future to remove residual UV filters from environmentally-friendly soils to remove hard-to-decompose substances in the environment.

碩士
國立交通大學
電信工程研究所
99
Typically, the leaky wave antenna is fed by a complex feeding structure to excite the higher order mode and it exhibits forward angle scanning characteristic. The composite right/left handed (CRLH) leaky wave antenna exhibits continuous backward-to-forward angle scanning characteristic and the broadside radiation capability. Moreover, it can be fed by a very simple and efficient (small and broadband) feeding structure. Mushroom structure exhibits unique electromagnetic properties, and it can be used for one kind of the 2-dimentional metamaterials. In the thesis, we use the eigenmode to analysis the dispersion diagram of the mushroom structure and we observe the electric field and magnetic field of each mode to understand characteristics of the mushroom structure. We acquire the radiation region from the dispersion diagram, and the mushroom structure can be represented by an equivalent circuit model. One of the mode solved by eigenmode method shows that this structure is unable to exhibit backward leaky wave; the circuit model constructed, however, predicts that the structure should be able to support backward leaky wave. We verified this contradiction by measuring the actual structure. First, we use single microstrip line to feed the composite right/left handed leaky wave antenna composed by the mushroom structure. However, the antenna gain is lower in the left-handed leaky wave region than in the right-handed leaky wave region. Thus, we use a 2-way power divider to feed the antenna. The bandwidth of the antenna fed by 2-way divider is from 5.6 GHz to 9.35 GHz, and the angle of radiation rotates clockwise from -51 to 69 degree. The antenna gain is over 10 dBi within the bandwidth. In addition, we acquire the angle of radiation of the main beam by far-field measurement, so we can calculate the phase constant.

碩士
國立交通大學
電信工程研究所
102
In this paper, a microstrip patch antenna loaded with mushroom-like structure is presented. Mushroom-like structure is composed of a mushroom with two L-shaped slots. This composite right/left-handed (CRLH) structure can excite zeroth-order res-onance (ZOR) and first-positive-order resonance (FPOR). This antenna has dual mode pattern includes monopolar and patch-like radiation pattern. ZOR frequency generates monopolar radiation pattern, FPOR generates patch-like radiation pattern. Generally speaking, FPOR frequency and first- negative-order resonance (FNOR) are restricted to the number of unit cells. Once the number of the unit cells is decided, the FPOR frequency cannot be adjusted. The first order resonance frequency of our proposed antenna can be adjusted by varying spacing of two L-shaped slot so that the FPOR frequency can be reduced. Proposed antenna consists of mushroom-like structure in the bottom and top mi-crostrip patch, and air layer is between twos substrates. As microstrip patch stacked with mushroom-like structure, antenna revealed triple-band behavior, and the third frequency can be determined by top patch size. Antenna is working in 2.31 GHz, 2.92 GHz, 3.7GHz.

Tese de mestrado. Biologia (Biologia Humana e Ambiente). Universidade de Lisboa, Faculdade de Ciências, 2012
Hoje em dia, devido ao prolongamento da esperança média de vida, bem como o aumento das agressões sofridas pelo meio ambiente envolvente, o cancro é uma das doenças mais proeminentes no seculo XXI. Neoplasmas, conjuntos anormais de células cromossomicamente mutadas, levam ao crescimento relativamente autónomo de tecido danificado. As causas destas mutações variam entre internas - condições imunitárias, hormonais, heritabilidade - ou externas - como tabagismo, exposição a radiação ou agentes químicos infecciosos (Zaidman et al., 2005). As células neoplásicas caracterizam-se pela perda das suas funções especializadas e ganham por sua vez características biológicas tais como a produção de sinais de crescimento, inibição da apoptose, elevado potencial replicativo e a capacidade de invadir tecidos circundantes (Zaidman et al., 2005; Zong et al.,, 2012). A esta metastização dos tecidos está associada 90% da responsabilidade da morte por cancro (Zong et al.,, 2012). Apesar da eficiência dos fármacos gerados até hoje no combate ao cancro, o seu efeito tóxico causa na maioria dos casos um decréscimo acentuado na qualidade de vida dos doentes. Assim, torna-se importante descobrir novos agentes anti cancerigenos com efeitos tóxicos mais baixos (Cao et al., 2010) que possam ser utilizados em terapias complementares. Virámo-nos então de novo para as tradições mais antigas da medicina Chinesa e Mediterrânea. Polissacáridos são biopolímeros constituídos por várias unidades de monossacáridos ligados através de ligações glicosidícas, que podem apresentar diferentes conformações, não só nas suas unidades constituintes, mas também na conformação das suas ligações, atribuindo-lhes assim diversas funções biológicas. Estes estão presentes desde o armazenamento de energia (amido), formação de componentes estruturais (celulose), participantes na comunicação célula a célula, possuindo ainda papéis importantes no sistema imunitário, na fertilização, desenvolvimento, agregação plaquetária e prevenção de patogéneses (Zong et al., 2012). Desde a medicina ancestral chinesa que os cogumelos têm sido reportados como tendo propriedades imunomoduladoras - actuando como modificadores da resposta biológica - e possuindo propriedades antivirais, antimicrobiais, circulatórias e nas ultimas décadas anti cancerigenas. (Wasser, 2010). A descoberta do Lentinian, um polissacárido com propriedades anti cancerígenas, despoletou nos anos 60 a pesquisa que tem sido dedicada ate hoje ao campo dos polissacáridos (Cao et al., 2010). Ao longo do tempo, diversos polissacáridos produzidos por cogumelos, têm demonstrado largos efeitos inibidores em vários tipos de tumores como Sarcoma 180, carcinoma de Lewis e Yoshida Sarcoma (Zhang et al., 2007). Existem três mecanismos propostos de actuação destes polissacáridos: (1) actividade preventiva, (2) aumento da resposta imunitária do hospedeiro e (3) inibição directa quando em contacto com células cancerígenas (Zhang et al., 2007). Na sua maioria os cogumelos que possuem polissacáridos biologicamente activos pertencem ao grupo dos basidiomicetes contendo cerca de 700 espécies descritas (Wasser, 2002). O seu poder antioxidante também desempenha um papel fundamental contra o desenvolvimento cancerígeno. Anti oxidantes naturais encontrados em fruta, vegetais, cogumelos e outros recursos alimentares têm sido investigados para prevenção de doenças coronárias e outras como o cancro (Kozarski et al.,2012). Assim, na procura de compostos não tóxicos que possuíssem propriedades anticancerígenas investigou-se também um pouco dos efeitos que as folhas de oliveira possam ter no combate ao cancro. A oliveira (Olea europaea) é uma árvore da família das Oleaceae, (Zaid et al., 2012) com folhas alongadas, que se encontra ao longo de toda região Mediterrânea desde há mais de 7000 anos (Lalas et al., 2011). Ao longo da história as folhas de Olea europaea têm sido exploradas para a prevenção de hipertensão, carcinogénese, diabetes, arteroesclerose bem como outras doenças do foro mais comum (Bouallagui et al., 2011; Zaid et al., 2012). A folha de oliveira foi ainda utilizada ao longo da história como um meio de combate á malária (Lee et al., 2009) A incidência de cancro ao longo do mediterrâneo tem sido das mais baixas quando comparada com outras partes do mundo, principalmente quando nos referimos ao cancro da mama, do endométrio, próstata e leucemia. (Bouallagui et al., 2011; Zaid et al., 2012) Assim reunimos neste trabalho, o estudo dos efeitos de algumas espécies conhecidas (e outras menos conhecidas) de cogumelos, com o estudo dos efeitos de algumas variedades de oliveiras nacionais, na inibição do crescimento de células tumorais. Para isso, foram efectuados vários doseamentos de polissacáridos, proteínas, açucares e compostos fenólicos e foram aplicadas diversas amostras das espécies amostradas a quatro linhas de carcinoma humano: HeLa, carcinoma cervical, A549 carcinoma pulmonar; A431 carcinoma epidermóide e OE21 carcinoma esófagofaringeal. De entre os variados resultados obtidos, pudemos concluir que a aplicação de extra polissacáridos da estirpe Ganoderma carnosum revelou um alto teor inibitório, e que os intra polissacáridos de todas as estirpes amostradas mostraram uma capacidade antioxidativa acima de 50%. As folhas de oliveira por sua vez, foram também capazes de mostrar um potencial anti proliferativo das células tumorais bastante satisfatório. Acreditamos que nestes compostos podem estar presentes as condições para a sua aplicação como terapia complementar às terapias anticancerígenas existentes.
Cancer is widely known nowadays as the disease of the century. Not only by the number of infected people worldwide but also by the amount of research dedicated to this field and its increasing success rate. Neoplasms, which are abnormal masses or colonies of cells produced by a relatively autonomous new growth of tissue, arise normally by the clonal expansion of a single cell that has undergone mutation in the chromosomal DNA, caused by a chemical, physical or biological agent (Zaidman et al., 2005). One of the most characterizing features of cancers, besides it’s ability to resist treatment, it’s his ability to metastasize and invade surrounding tissues, causing 90% of cancer deaths ( Zong et al., 2012). Although the efficiency of chemotherapy for the majority of cancers has improved over the last three decades, the drugs used in this treatment, contain high toxic effects that cause severe reduction in quality of life. Therefore, it is important to develop novel potent, but low toxic anti-cancer reagents, including natural products (Cao et al., 2010). The desire to find this new chemical drug as lead us to look back in time and to report to times of ancient Chinese and Mediterranean medicine, in order to encounter natural products that can in the future be developed as efficient co-chemotherapeutical agents. During this work we will test mushroom polysaccharides as well as olive leaves phenolic compounds, in human carcinoma cell lines, in order to discover more about their anti-tumor effects. Amongst our findings, extrapolysaccharides from Ganoderma lucidum were the most promising and all intrapolysaccharides were found to have a 50% rate of antioxidative power. Olive leaves were also very promising in the inibithion of human carcinoma cells in vitro.

Dissertação de Mestrado Integrado em Engenharia Química apresentada à Faculdade de Ciências e Tecnologia
Presently, the rapid rate of growth of the affected population with premature aging and numerous diseases is evident being cancer and neurodegenerative diseases, such as Alzheimer's, Parkinson's and amyotrophic lateral sclerosis, the most prevalent and requiring more attention. One of the main causes for the onset of these diseases is the uncontrolled production of reactive oxygen species (ROS), derived from an excessive production of free radicals. In order to reduce oxidative damage and considering the restrictions on the use of synthetic antioxidants, there is an increasing demand for natural products that contain a high antioxidant potential.Among the existing natural products, mushrooms have been the subject of countless studies, due to their beneficial effects on health in addition to their nutritional properties. These are fungi quite rich in bioactive compounds, such as polysaccharides, phenolic compounds, vitamins and secondary metabolites. Some of the referred compounds have numerous antioxidant properties that are very beneficial to the organism. This master's thesis project is focused on the study of the Tricholoma equestre mushroom species, which has not been much studied until now. The scope of the work was to investigate the effect of polysaccharides and of phenolic compounds, extracted from Tricholoma equestre species, on the production of ROS in neuronal cells. The experiments were performed in brain slices from Wistar rats, at the mossy fiber synapses from CA3 hippocampal area. Polysaccharides and phenolic compounds extracts obtained from the Tricholoma mushroom, were used to prepare different solutions with concentrations of 0.1, 0.5 and 1 g.L-1, by adding the extracts to the artificial cerebrospinal fluid (ACSF), which has the composition of the neuronal extracellular medium. The aim of this part of the work was to study the effect of those compounds on neuronal ROS production using the fluorescent ROS indicator H2DCFDA.The results show that the lower polysaccharides concentrations (0.1 and 0.5 g.L-1) did not cause significant ROS changes while the higher dose (1 g.L-1) led to a decrease of the ROS signals of about 2.7 % with respect to the baseline. The experiments carried out with the phenolic compounds solutions gave more accentuated changes that were opposite to the previous one. The larger increases in ROS production were observed for the concentrations of 0.1 and 1 g.L-1, having the fluorescence signals amplitudes of about 18 % and 14 %, respectively, while for 0.5 g.L-1 the enhancement was approximately 6 %. These facts indicate that the phenolic compounds have a peculiar concentration dependent effect, occurring the larger ROS changes for the lower concentration, 0.1 g. L-1, whose signals have a similar profile to that of the higher concentration, 1 g. L-1.The last group of experiments was made with the synthetic compound, 4-hydroxybenzoic acid (1 g.L-1), which is the only identified phenolic compound of the studied mushroom species. The results indicate that this acid causes similar changes to those observed for the same concentration of the phenolics extract, with a maximum value of about 14 % upon the removal of the acid.In conclusion, and due to the obtained results, it is undeniable that mushrooms have antioxidant properties that affect the performance of cellular metabolism, acting on ROS signals and triggering different responses of the organism to these stimuli.
Atualmente, é evidente a rápida taxa de crescimento da população afetada com envelhecimento prematuro e por numerosas doenças como o cancro e doenças neurodegenerativas, tais como as doenças de Alzheimer, de Parkinson e a esclerose lateral amiotrópica, que são as mais frequentes e que requerem mais atenção. Uma das causas principais para o início destas doenças é a produção descontrolada de espécies reativas de oxigénio (ROS), derivadas da produção excessiva de radicais livres. De forma a reduzir danos oxidativos e considerando a restrição no uso de antioxidantes sintéticos, verifica-se uma procura crescente de produtos naturais com um elevado potencial antioxidante. Entre os produtos naturais existentes os cogumelos têm sido objeto de inúmeros estudos devido aos seus efeitos benéficos na saúde para além das suas propriedades nutricionais. Estes são fungos muito ricos em compostos bioativos, tais como polissacarídeos, compostos fenólicos, vitaminas e metabolitos secundários. Alguns dos compostos referidos tem muitas propriedades antioxidantes que são muito benéficas para o organismo. Este projeto de tese de mestrado foca-se no estudo da espécie de cogumelos Tricholoma equestre que, até à data, não tem sido muito estudado. O objetivo do estudo era investigar o efeito de polissacarídeos e de compostos fenólicos, extraídos da espécie Tricholoma equestre, na produção de ROS em células neuronais. As experiências foram realizadas em fatias cerebrais de ratos Wistar nas sinapses das fibras musgosas da área CA3 do hipocampo. Extratos de polisacarídeos e de compostos fenólicos, obtidos do cogumelo Tricholoma, foram usados para preparar diferentes concentrações de 0.1, 0.5 e 1 g.L-1, adicionando os extratos ao fluído cerebroespinal artificial (ACSF), que tem a composição do meio neuronal extracelular. O objetivo desta parte do trabalho era estudar o efeito daqueles compostos na produção de ROS neuronal usando o indicador de ROS fluorescente H2DCFDA.Os resultados mostram que as concentrações mais baixa de polissacarídeos (0.1 and 0.5 g.L-1) não causaram variações significativas de ROS enquanto que a dose mais alta (1 g.L-1) levou a uma diminuição dos sinais de ROS de cerca de 2.7 % em relação à linha base. As experiências realizadas com as soluções de compostos fenólicos originaram variações mais acentuadas que eram opostas à anterior. Os maiores aumentos na produção de ROS foram observados para as concentrações de 0.1 e 1 g.L-1, tendo os sinais de fluorescência respetivamente amplitudes de cerca de 18 % e 14 %, enquanto que para 0.5 g.L-1 o aumento era aproximadamente 6 %. Estes factos indicam que os compostos fenólicos têm um efeito dependente da concentração que é peculiar, ocorrendo as maiores variações de ROS para a menor concentração, 0.1 g.L-1, cujos sinais têm um perfil semelhante aos da concentração mais elevada, 1 g. L-1. O último grupo de experiências foi feito com o composto sintético 4-hydroxybenzoic acid (1 g.L-1), que é o único composto fenólico identificado na espécie de cogumelos estudada. Os resultados indicam que este ácido causa variações de ROS semelhantes às que foram observadas para a mesma concentração do extrato de fenólicos, com um valor máximo de cerca de 14 % após a retirada do ácido.Em conclusão, e pelos resultados obtidos, é inegável que os cogumelos possuem propriedades antioxidantes que afetam o desempenho de metabolismo celular, atuando nos sinais de ROS e desencadeando diferentes respostas do organismo a estes estímulos.
Universidade de Coimbra - Compra de materiais, reagentes e equipamentos referentes a realização experimental da dissertação de mestrado.

碩士
國立交通大學
電信工程研究所
102
In Thesis, the new type of compact mushroom structure had been proposed. The original equivalent circuit transmission line model tells that the electromagnetic bandgap (EBG) range can be affected by its equivalent capacitor and inductor. So this thesis propose the simple way to shift down the EBG range by inserting lunped element, instead of complex design, such as interdigital capacitor and meander line. Also we use three methods to analyze the compact mushroom structure, dispersion diagram, S-parameters measurement, and modified equivalent circuit model. Three of them have different advantage for analyzing. Dispersion diagram provides the complete analysis for compact mushroom structures, S-parameter measurement is the only way to observe the effects of EBG practically, and modified equivalent circuit model have the quicker way to find out the bandgap range. In addition, patch antenna and CRLH small antenna are designed to combine with compact mushroom structure, to prove that the radiation efficiency of both antenna can be improved by compact mushroom structure.

碩士
國立交通大學
電信工程研究所
103
In this thesis, a compact tri-band and dual-polarized antenna, using a combination of mushroom-like structure and square-ring patch antenna is presented. This antenna can excite one linear polarization and two circular polarization. Mushroom-like structure is evolved from conventional mushroom structure for circular polarization by slotting slits along the diagonal direction of patch. This mushroom structure can be equivalent to composite right/left-handed transmission line model, and can realize dual-band and dual-polarized antenna that provides linear polarization at zeroth-order resonance (ZOR) and circular polarization at first-positive-order resonance (FPOR). Circular polarization is excited by using truncated corner patch on mushroom structure at FPOR. In order to achieve frequency tunability, the ZOR frequency can be adjusted by changing the left-handed inductance value or the right-handed capacitance value. Furthermore, the FPOR frequency can be adjusted by slotting different length of slits to change different current path, and the ZOR frequency is almost fixed. For realizing the third resonant frequency, the mushroom-like structure antenna combines with square-ring patch antenna by placing mushroom-like structure inside square-ring patch antenna in this thesis. Conventional microstrip patch antenna operating at fundamental mode is equal to half-wavelength resonance, but square-ring patch antenna that can decrease resonant frequency by extending current path is able to achieve antenna miniaturization, also uses truncated corner ring to excite circular polarization. Taking advantage of coaxial feed and coupling-strip feeds directly signal to two antennas, so impedance matching is easy to reach. The advantage of this tri-band and dual-polarized antenna is that all of the antennas are realized on upper metal of the same substrate, and only need single signal feeding to reach tri-band and dual-polarization. Finally, this antenna operates at 1.9GHz for ZOR, 2.45GHz for fundamental mode of square-ring patch antenna and 3.5GHz for FPOR.

A 24-hour fermentation of the optimized synbiotic composed of B. longum and EPR was performed to give a cell-free fermentation broth (S24). S24 was co-cultured with two colon cancer cell lines (Caco-2 and SW620) and normal colon cells (FHC). S24 significantly inhibited cell proliferation for both colon cancer cells but promoted FHC cell growth by 10-25% as shown by MTT and BrdU arrays. Primary DNA damage analysis by alkaline comet assay showed S24 caused DNA damage to a comparable extent as the positive control of 10 mM H2O2 (treated for 1 hour) for both cancer cells. Dynamic analysis on DNA damage-associated DNA repair showed the two colon cancer cells had different response pattern to S24. Flow cytometric analysis showed that both Caco-2 and SW620 when treated with S24 (IC 50=3.66 mM of acetate) were arrested initially at G2/M and subsequently at S phase accompanied with large sub-G1 peaks. Dual staining with PI/AnnexinV further proved the appearance of apoptosis. Live cell imaging analysis on Caco-2 cells treated with S24 showed the following events: mitochondria were rapidly destroyed within the first two-hour treatment, the cells bubbled and the nucleus condensed after the mitochondrial had shrunken, followed by apoptosis.
Despite active research on synbiotic on anti-carcinogenesis of colon cancer by synbiotics, the underlying mechanism still remains unclear. This study investigated a novel synbiotic composed of non-digestible carbohydrates (NDCs) extracted from mushroom sclerotia as prebiotics and Bifidobacteria as probiotics. Preliminary results on incubation of two probiotics ( Bifidobacterium longum and Lactobacillus brevis) and one pathogenic bacterium (Clostridium celatum) separately with 3 NDCs extracted from mushroom sclerotia [Poria cocos (PC), Polyporus rhinocerus (PR) and Pleurotus tuber-regium (PT)] indicated that the growth of B. longum and L. brevis was stimulated more preferentially than C. celatum after 72-hour fermentation. The short-chain fatty acid (SCFA) profile was dominated by acetate (> 98% of total SCFAs) with very little butyrate (< 2.0% of total SCFAs) and the organic matter disappearance (OMD) during fermentation was consistent with the bacterial growth. Among the synbiotic combinations, NDC from PR and B. longum gave the largest amount of acetate (2.47+/-0.232 mmol/g of organic matter disappearance).
Results obtained from human pathway finder RT2 Profiler(TM) PCR Array indicated that S24 could modulate the proliferation of colon cancer cells mainly by various pathways such as cell cycle and DNA damage repair, apoptosis and cell senescence, etc. In SW620 cells, PCR Array of Human Cell Cycle further revealed that the modulated genes mainly belonged to the gene cluster of S phase and DNA replication as well as G2 and G2/M transition. While for Caco-2 cells, the cell-cycle modulated genes mainly belonged to the cluster of G2 and G2/M transition. Immuno-blotting on the pivotal upstream regulators showed that phosphorylation of ATM at Serine 1981 was significantly increased in both cancer cells. Site-specific phosphorylation of pRB was decreased and phosphorylation of Chk1 was increased in both cancer cells while Chk2 were increased in SW620 cells. Cdc25A was phosphorylated at serine17 in both cancer cells. It can be proposed that the blockage of DNA synthesis or DNA damage was due to the down-regulation of some pivotal DNA replication related proteins such as RPA3, PCNA and MCMs, detected by ATM-Chk1/Chk2-Cdc25A pathway. This would cause the prolonged staying of cells at the G1/S checkpoint which further moved on to S phase arrest for SW620 cells. Moreover the sharply up-regulated p21, an important inhibitor of Cdk2 would further hinder the cells passing the G1/S checkpoint in SW620 cells.
The tumor suppressor p53 was detected phosphorylated at various sites in SW620 but not in Caco-2 cells. In SW620 cells, G2/M arrest was caused by the inhibition of CDK1/CDC2 due to increased expression of GADD45A and p21 and phosphorylation of Cdc25A, while for Caco-2, the G2/M arrest was caused by degradation of Cdc25A due to the absence of p53-activated GADD45A and p21 expression as shown in the pathway finder results. Some apoptosis-related proteins of Bax, Apaf-1 and PARP were modulated as shown by immuno-blotting in both colon cancer cells. (Abstract shortened by UMI.)
Gao, Shane.
Adviser: Peter Chi-Keung Cheung.
Source: Dissertation Abstracts International, Volume: 72-11, Section: B, page: .
Thesis (Ph.D.)--Chinese University of Hong Kong, 2009.
Includes bibliographical references (leaves 55-94).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.