O câncer é a segunda causa de morte da população mundial e constitui-se em sério problema de saúde pública no mundo. Seu tratamento acarreta além de elevados gastos, diversos efeitos colaterais aos pacientes. Dessa forma, alternativas que possam minimizar esses fatos são extremamente viáveis. Diversos estudos já comprovaram o efeito sinérgico dos isotiocianatos e do selênio e a sua contribuição como agentes protetores contra danos oxidativos e aumentam a expressão de enzimas responsáveis pela desintoxicação de compostos cancerígenos. Com base nisso, o objetivo desta pesquisa foi avaliar a influência da suplementação com selênio, durante o processo de maturação do brócolis, em parâmetros das atividades antioxidantes e antiproliferativo. No presente estudo foi constatado que a biofortificação do brócolis com selênio é possível uma vez que, as amostras tratadas foram capazes de absorver este mineral. Além disso, a biofortificação acarretou o aumento tanto da atividade antioxidante como dos compostos fenólicos em todas as amostras testadas. A avaliação do potencial antiproliferativo do brócolis demonstrou que os extratos brutos diclorometânicos apresentaram tal potencial. Os extratos brutos diclorometânicos das mudas de brócolis biofortificadas com selênio se destacaram perante os demais extratos quanto a atividade antiproliferativa. Concluindo, a biofortificação com selênio constitui-se uma técnica viável para o aumento do potencial antioxidante do brócolis e os estudos iniciais sobre a atividade antiproliferativa demonstram que as mudas de brócolis biofortificadas com selênio são as amostras mais promissoras para realização de estudos futuro. / Worldwide cancer is the second leading cause of death and constitutes a serious public health problem. The traditional treatment beyond expensive has several side effect, thus alternatives that can minimize these realities are extremely viable. Isothiocyanates and selenium have synergic effect, contributes as protective agents against oxidative damage and increase the expression of detoxification enzymes responsible for eliminate carcinogenic compounds. Therefore, the aim of this research was to evaluate the influence of selenium supplementation during the broccoli maturation in antioxidant and antiproliferative parameters. In the present study it was found that broccoli biofortification with selenium is possible since plants were able to absorb this mineral. Furthermore, an increase of both antioxidant activity and phenolic compounds was observed in all samples supplemented. All dichloromethanic crude extracts showed better antiproliferative activity when compared with ethanolic extracts being seedlings supplemented with selenium the best antiproliferative agent among the extracts tested. In conclusion, biofortification with selenium constitutes a viable technique for increasing the antioxidant potential of broccoli. Initial studies with antiproliferative activity showed that the broccoli seedlings biofortified with selenium are the most promising samples for future studies.

Submitted by RAPHAEL LEONE DA CRUZ FERREIRA null (rapha.leone@hotmail.com) on 2016-10-10T20:41:26Z No. of bitstreams: 1 DISSERTAÇÃO DE MESTRADO.pdf: 1223687 bytes, checksum: 7eb478241afa939e0b2a76387a523f55 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-10-17T13:33:25Z (GMT) No. of bitstreams: 1 ferreira_rlc_me_jabo.pdf: 1223687 bytes, checksum: 7eb478241afa939e0b2a76387a523f55 (MD5) / Made available in DSpace on 2016-10-17T13:33:25Z (GMT). No. of bitstreams: 1 ferreira_rlc_me_jabo.pdf: 1223687 bytes, checksum: 7eb478241afa939e0b2a76387a523f55 (MD5) Previous issue date: 2016-09-08 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / As informações sobre fontes de selênio (Se) em alface são incipientes na literatura, sobretudo, com relação ao limite entre biofortificação e toxicidade. Assim, o objetivo deste estudo foi determinar níveis críticos de Se na solução nutritiva e foliar, e a melhor fonte que aumente a biofortificação da alface hidropônica sem causar toxicidade. O delineamento experimental utilizado foi inteiramente casualizado representado por duas fontes de selênio, selenito de sódio (Na2SeO3) e selenato de sódio (Na2SeO4), dez concentrações de selênio (0; 2; 4; 6; 8; 16; 32; 64; 96; 128 μM) com quatro repetições por tratamento, totalizando 80 unidades experimentais. As concentrações adequadas estão entre 5,6 e 16 μM de selênio na forma selenato em solução nutritiva e teores foliares adequados de 10,2 a 41,8 mg kg-1 de selênio. / Information on sources of selenium (Se) in lettuce are scarce in the literature, especially with respect to the boundary between Biofortification and toxicity. The objective of this study was to determine critical levels of Se in the nutrient solution and leaf and the best source to increase biofortification of hydroponic lettuce without causing toxicity. The experimental design was completely randomized represented by two sources of selenium, sodium selenite (Na2SeO3) and sodium selenate (Na2SeO4) ten selenium concentrations (0. 2. 4. 6. 8. 16. 32. 64. 96. 128 M) with four replicates per treatment, totaling 80 experimental units. The concentrations are between 5.6 and 16 M selenium selenate form the nutrient solution and appropriate foliar 10.2 to 41.8 mg kg-1 of selenium. / CNPq: 132361/20159

O câncer é a segunda causa de morte da população mundial e constitui-se em sério problema de saúde pública no mundo. Seu tratamento acarreta além de elevados gastos, diversos efeitos colaterais aos pacientes. Dessa forma, alternativas que possam minimizar esses fatos são extremamente viáveis. Diversos estudos já comprovaram o efeito sinérgico dos isotiocianatos e do selênio e a sua contribuição como agentes protetores contra danos oxidativos e aumentam a expressão de enzimas responsáveis pela desintoxicação de compostos cancerígenos. Com base nisso, o objetivo desta pesquisa foi avaliar a influência da suplementação com selênio, durante o processo de maturação do brócolis, em parâmetros das atividades antioxidantes e antiproliferativo. No presente estudo foi constatado que a biofortificação do brócolis com selênio é possível uma vez que, as amostras tratadas foram capazes de absorver este mineral. Além disso, a biofortificação acarretou o aumento tanto da atividade antioxidante como dos compostos fenólicos em todas as amostras testadas. A avaliação do potencial antiproliferativo do brócolis demonstrou que os extratos brutos diclorometânicos apresentaram tal potencial. Os extratos brutos diclorometânicos das mudas de brócolis biofortificadas com selênio se destacaram perante os demais extratos quanto a atividade antiproliferativa. Concluindo, a biofortificação com selênio constitui-se uma técnica viável para o aumento do potencial antioxidante do brócolis e os estudos iniciais sobre a atividade antiproliferativa demonstram que as mudas de brócolis biofortificadas com selênio são as amostras mais promissoras para realização de estudos futuro. / Worldwide cancer is the second leading cause of death and constitutes a serious public health problem. The traditional treatment beyond expensive has several side effect, thus alternatives that can minimize these realities are extremely viable. Isothiocyanates and selenium have synergic effect, contributes as protective agents against oxidative damage and increase the expression of detoxification enzymes responsible for eliminate carcinogenic compounds. Therefore, the aim of this research was to evaluate the influence of selenium supplementation during the broccoli maturation in antioxidant and antiproliferative parameters. In the present study it was found that broccoli biofortification with selenium is possible since plants were able to absorb this mineral. Furthermore, an increase of both antioxidant activity and phenolic compounds was observed in all samples supplemented. All dichloromethanic crude extracts showed better antiproliferative activity when compared with ethanolic extracts being seedlings supplemented with selenium the best antiproliferative agent among the extracts tested. In conclusion, biofortification with selenium constitutes a viable technique for increasing the antioxidant potential of broccoli. Initial studies with antiproliferative activity showed that the broccoli seedlings biofortified with selenium are the most promising samples for future studies.

No description available.

Precipitation over coal fly ash surfaces raises concerns about potential downstream impacts of dissolved selenium (Se) on aquatic ecosystems. Detention ponds hold runoff until aqueous Se concentrations meet water quality regulations: within 10 days, 90% of runoff must be discharged and cannot exceed 11.8 μg Se L-1 (ppb). Constructed wetland (CW) phytoremediation is a potential treatment option to meet these regulations. This research was conducted to 1.) Assess the potential of native southeastern aquatic macrophytes for Se tolerance and removal efficiency compared to unplanted (UNP) detention ponds; 2.) Evaluate seasonal influence on Se removal in CWs over four, weekly flood-discharge cycles; and 3.) Determine the effect of temperature on aqueous Se removal. Monoculture CW microcosms (110 L) were flooded six days with 500 or 1000 ppb Se, as selenite (SeO32-) or selenate (SeO42-) evaluating five plant species for Se tolerance. Seasonal evaluations were conducted with cattail (Typha angustifolia L.; CAT) and duckweed (Lemna minor L.; DWD) over four weekly flood-discharge cycles at 16 or 32 ppb SeO42--Se. Cattail, DWD, or UNP volatilization chambers (VCs) flooded with a 3 L solution at 35 ppb SeO42--Se were evaluated under 12 h photoperiod at either 20 or 30°C. All experiments contained a zero Se control. Water, plant, and soil total Se concentrations were determined by inductively coupled plasma-mass spectroscopy (ICP-MS) and analyzed with PROC GLM (SAS EG 7.1) at α=0.05. Cattail and DWD were selected for further evaluation with SeO42--Se. The greatest aqueous Se removal was observed in the summer (73%), followed by the fall (42%) and spring (41%), compared to winter (18%). Temperature was strongly correlated with Se removal (0.65, P<0.0001). Except for summer, after two flood-discharge cycles, CAT and DWD improved aqueous Se removal compared to UNP controls. Volatilization chamber data indicated greater Se removal at 30 (69%) compared to 20°C (54%). At 30°C, DWD decreased aqueous Se concentrations below 11.8 ppb Se within 10 days. Given their efficacy and abundance, CAT and DWD are deemed suitable species for phytoremediation in CWs supplied with Se-impacted waters.

No description available.

The objective of this study was to evaluate the influence of biofortification with selenium in the bioactive compounds of broccoli; to test their toxicity and mutagenicity in vivo to prove their safety for consumption; to develop microparticles by the spray dryer technique and to evaluate its physicochemical properties. The broccoli seedlings were biofortified through the addition of 2 mL of 50 &mu;M sodium selenate solution 15 days after sowing (DAS), and collected 30 DAS. The samples were analyzed by different methods for the quantification of selenium (ICP-MS, benchtop and handheld EDXRF). The energy-dispersive X-ray fluorescence spectrometry made either in bench or in a portable equipment presented a good performance for the quantification of selenium, together with low cost and less analysis time. The selenium biofortification technique increased the concentration of this micronutrient in broccoli seedlings. In addition, it also allowed a significant increase in bioactive compounds, such as chlorophyll (12%), phenolic (26%) and flavonoid (19%) compounds, vitamin C (32%) and sulforaphane (12%), the main bioactive compound with anticarcinogenic potential. The efficiency of biofortification with selenium in the culture was verified, and toxicity and mutagenicity studies were conducted in female and male mice treated with different doses of selenium-biofortified broccoli seedlings. After 40 consecutive days of treatment, broccoli seedlings with selenium at doses of 45 and 70 &mu;g Se/kg body weight resulted in significant changes in the relative weights of vital (liver, spleen and kidneys) and reproductive organs (ovary/testis), with the presence of histological alterations in the kidneys and spleen in mice of both sexes. In male mice, the dose of 70 &mu;g Se/kg body weight increased the frequency of micronucleated polychromatic erythrocytes. In females this increase was not observed, demonstrating greater sensitivity of males to the treatment. Finally, microparticles of ethanolic extracts of broccoli seedlings with and without selenium were developed through the spray drying microencapsulation technique. The samples presented low moisture (4-5%) and hygroscopicity (11-12 g of water absorbed 100/g of microparticle) and high solubility (99%). However, there was a significant reduction in the content of phenolic compounds and antioxidant activity after the microencapsulation process. The results of this study demonstrate the importance of biofortification of selenium as a strategy to increase the content of bioactive compounds in broccoli and that the dose of 15 &mu;g Se/kg body weight had no significant toxic or mutagenic effects. In addition, microencapsulation has been shown to allow the elaboration of samples with good technological characteristics however, optimization studies should be conducted to increase the retention power of bioactive compounds and their antioxidant activity. / Os objetivos desse estudo foram avaliar a influência da biofortificação com selênio nos compostos bioativos de brócolis; testar sua toxicidade e mutagenicidade in vivo como forma de comprovar sua segurança para consumo; desenvolver micropartículas pela técnica de spray dryer e avaliar suas propriedades físico-químicas. As mudas de brócolis foram biofortificadas pela adição de 2 mL de solução de 50 &mu;M de selenato de sódio 15 dias de após a semeadura (DAS) e coletadas aos 30 DAS. As amostras foram analisadas por diferentes métodos para a quantificação de selênio (ICP-MS, XRF portátio e EDXRF de bancada). O método de fluorescência de Raios X por dispersão em energia feito em equipamentos de bancada ou portátil apresentou desempenho adequado para quantificação de selênio aliado à baixo custo e menor tempo de análise. A técnica de biofortificação com selênio aumentou a concentração desse micronutriente nas mudas de brócolis. Além disso, também proporcionou aumento significativo nos compostos bioativos, tais como, clorofila (12%), compostos fenólicos (26%) e flavonoides (19%), vitamina C (32%) e sulforafano (12%), principal composto bioativo com potencial anticarcinogênico do brócolis. Verificada a eficiência da biofortificação com selênio, conduziu-se estudos de toxicidade e mutagenicidade em camundongos fêmeas e machos tratados com diferentes doses de mudas de brócolis biofortificadas com selênio (15, 45 e 70 &mu;g Se/kg peso corporal). Após 40 dias consecutivos de tratamento, observou-se que os animais tratados com mudas de brócolis com selênio nas doses de 45 e 70 &mu;g Se/kg peso corporal provocaram significativas alterações nos pesos relativos dos órgãos vitais (fígado, baço e rins) e reprodutivos (ovário/testículos), com presença de alterações histológicas nos rins e no baço, em camundongos de ambos os sexos. Nos camundongos machos, a dose de 70 &mu;g Se/kg peso corporal provocou aumento da frequência de eritrócitos policromáticos micronucleados. Nas fêmeas este aumento não foi observado, demonstrando maior sensibilidade dos machos ao tratamento. Por fim, através da técnica de microencapsulação por spray dryer desenvolveu-se micropartículas a base de extratos etanólicos de mudas de brócolis com e sem selênio. As amostras apresentaram baixa umidade (4-5%) e higroscopicidade (11-12 g de água absorvida 100-1 g de micropartícula) e alta solubilidade (99%). No entanto, houve uma redução significativa no teor de compostos fenólicos e atividade antioxidante após o processo. Os resultados desse estudo demonstram a importância da biofortificação do selênio como uma estratégia para aumentar o teor de compostos bioativos no brócolis. Com relação à toxicidade, a dose de 15 &mu;g Se/kg peso corporal não apresentou efeitos tóxicos significativos em machos e fêmeas, porém apresentou pontencial mutagênico em machos. Além disso, comprovou-se que a microencapsulação possibilitou a elaboração de amostras com boas características tecnológicas, porém, estudos de otimização devem ser conduzidos a fim de aumentar o poder de retenção dos compostos bioativos e atividade antioxidante.

Open-framework inorganic and inorganic-organic hybrid materials constitute an important area of study in materials chemistry, because of their potential applications in areas such as sorption and catalysis. Besides aluminosilicate zeolites, the metal phosphates and carboxylates constitute large families of open-framework structures. The possibility of building open architectures with the sulfate and selenate anions as the basic building units has been explored in this thesis. Investigations of a variety of open-framework metal sulfates and selenates, as well as a family of jarosites of different transition metals are presented. More importantly, studies directed towards the synthesis and understanding of the magnetic properties of various Kagome compounds formed by the transition metal ions is discussed at length. After providing an introduction to inorganic open-framework compounds (Part 1), the thesis presents the results of the investigations of various transition and rare earth metal sulfates with diverse structures and dimensionalities in Part 2. Some of these compounds show interesting properties. For example, a two-dimensional Ni(II) sulfate exhibits ferrimagnetism whereas a three-dimensional Ni(II) sulfate with 10-membered channels is paramagnetic. A family of three-dimensional co-ordination polymers of Co(II) sulfate wherein the Co(II) sulfate layers are linked by diaminoalkanes of varying chain length has been synthesized and characterized. Organically-templated neodymium and thorium sulfates with layered and three-dimensional structure have also been prepared. The jarosite family of compounds with the Kagome structure is considered as an ideal model for studying frustrated magnetism. This type of materials, however, is difficult to prepare in a pure and highly crystalline form. We have synthesized analogues of the jarosite containing magnetic ions other than Fe3+ by solvothermal techniques and discussed them in Part 3. In particular, we have prepared and explored the magnetic properties of Mn2+(S = 5/2), Fe2+ (S = 2), Co2+(S = 3/2) and Ni2+ (S = 1) jarosites. Based on the results presented, it becomes clear that the magnetic properties vary with the spin of the transition metal ion. It appears that those Kagome compounds with transition metalions with non-integer spins show antiferromagnetic interactions and magnetic frustration while those with integer spins exhibit ferro/ferrimagnetic properties. A theoretical study has also supports this observation. We have been able to isolate for the first time 1,4-diazacubane as the part of the structure of the nickel Kagome compound. The possibility of building open architectures with the selenate anion as the basic building unit has been explored in Part 4. The results have been rewarding and an organically-templated three-dimensional lanthanum selenate with 12-membered channels has thus been obtained for the first time.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Selenium (Se) is an oligoelement crucial for various biological processes. Se has anti-inflammatory and antioxidant properties and plays a key role in brain development. Adenosine deaminase (ADA, EC 3.5.4.4) is a key enzyme in purine metabolism because it helps in the regulation of intracellular and extracellular levels of adenosine, an important nucleoside that acts in the neuromodulation of the immune and nervous systems. Oxidative stress occurs when there is an imbalance between the production of reactive species and their detoxification by systems that remove or repair the resulting damage. In this context, several Se compounds have been developed and studied, among them the 3-methyl-1-phenyl-2-(phenylseleno)oct-2-en-1-one (C12H2HOSe), which is an α, β-unsaturated ketone functionalized vinyl chalcogenide, and sodium selenate (Na2SeO4), the main inorganic form of Se found in animals and plants. The aim of this study was to compare the effects of C12H2HOSe (organic Se) and Na2SeO4 (inorganic Se) on the cell viability, lipoperoxidation and ADA activity in cortex slices of young rats. The results showed that only organic Se caused a reduction in ADA activity at the concentrations of 1, 10 and 30μM in the cerebral cortex slices of the rats tested. However, this result is not related to the oxidation of the thiol groups, since this compound did not alter the NP-SH and LDH levels. Both compounds did not affect the lipoperoxidation levels, although the organic Se was capable of sequestering NO radicals at the highest concentration tested. The inorganic Se protected the cerebral cortex slices against the sodium nitroprusside-induced damage and increased the NP-SH levels. The tests used to evaluate cell viability (LDH and MTT) suggest the maintenance of the cell integrity of the cortex slices exposed to Se compounds. Therefore, our results suggest that organic Se has immunomodulatory properties, due to the reduction in ADA activity, and acted in the maintenance of the cell integrity. In turn, the antioxidant activity of inorganic Se was reaffirmed. Hence, the results of this study paved the way to explore both Se compounds in the CNS. / O selênio (Se) é um oligoelemento essencial em vários processos biológicos. Possui propriedades antiinflamatórias, antioxidantes e desempenha um papel fundamental no desenvolvimento do cérebro. A adenosina deaminase (ADA, E.C. 3.5.4.4) é uma enzima chave no metabolismo das purinas, pois auxilia na regulação dos níveis intra e extracelulares de adenosina, um importante nucleosídeo que atua na neuromodulação dos sistemas nervoso e imune. O estresse oxidativo ocorre quando há desequilíbrio entre a produção de espécies reativas e sua desintoxicação através de sistemas que removem ou reparem os danos por elas causados. Nesse contexto, vários compostos de Se vêm sendo desenvolvidos e estudados, entre eles podemos citar o 3-metil-1-fenil-2-(fenilseleno)oct-2-en-1-um(C12H2HOSe),um organocalcogênio com uma cetona α, β- insaturada funcionando como um vinil calcogênio e o selenato de sódio (Na2SeO4), principal Se inorgânico encontrado em animais e plantas. Este estudo teve como objetivo comparar os efeitos do C12H2HOSe (Se orgânico) e Na2SeO4 (Se inorgânico) na viabilidade celular, parâmetros relacionados ao estresse oxidativo e atividade da ADA em fatias de córtex de ratos jovens. Os resultados demonstraram que apenas o Se orgânico provocou a diminuição na atividade da ADA nas concentrações de 1, 10 e 30μM nas fatias de córtex cerebral dos ratos testados. No entanto, esse resultado não tem relação com a oxidação de seus grupamentos tióis, já que esse composto não alterou os níveis de NP-SH e de LDH. Apesar de ambos os compostos não alteraram os níveis de lipoperoxidação, o Se orgânico apresentou capacidade de sequestrar radicais NO na maior concentração testada. Já o composto inorgânico de Se protegeu as fatias de córtex cerebral contra o dano induzido pelo nitroprussiato de sódio e aumentou os níveis de NP-SH. Os testes de viabilidade celular (LDH e MTT) sugerem a manutenção da integridade celular em fatias de córtex de ratos jovens quando expostos aos compostos de Se. Assim, nossos resultados sugerem que o Se orgânico apresenta propriedades imunomoduladoras por reduzir a atividade da ADA, além de atuar na manutenção da integridade celular. Já o Se inorgânico teve sua atividade antioxidante confirmada. Portanto, os resultados obtidos neste estudo destacam um caminho promissor a ser explorado por ambos os compostos no SNC.

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