Respuesta transcripcional de Populus a bifenilos policlorados: Aspectos aplicados a la fitorremediación

La degradación del suelo ha adquirido una magnitud preocupante. Los métodos tradicionales de descontaminación, son costosos e insuficientes. La fitorremediación representa una alternativa eficaz, de bajo coste, respetuosa con el medio ambiente, que además mejora las propiedades del suelo, si bien ha habido desarrollos relevantes en la última década. Desde el punto de vida científico, el reto principal es descifrar las rutas metabólicas implicadas en respuesta a contaminantes y comprender su regulación. Esta información es imprescindible si aspiramos a mejorar las capacidades naturales de algunas especies vegetales para remediar los suelos contaminados. Los estudios de esta Tesis se han centrado en Populus, el mejor modelo forestal disponible a raíz de la secuenciación de su genoma completo. Por otra parte, Populus tiene una gran capacidad natural para la degradación de contaminantes orgánicos, lo que explica su predominio en los programas forestales de fitorremediación que se desarrollan actualmente. Hemos elegido en concreto al híbrido Populus tremula x P. alba, por la facilidad con que se cultiva y su particular interés biotecnológico. La presente Tesis plantea un estudio comprehensivo de la respuesta molecular a bifenilos policlorados (PCBs), una familia de contaminantes orgánicos persistentes de particular relevancia a escala mundial. Se ha utilizado para ello una aproximación transcriptómica, basada en tecnología RNA-seq, para identificar los genes implicados en el metabolismo de los compuestos in planta y cuantificar sus niveles de activación en distintas situaciones controladas. La tesis pretende asimismo definir el control transcripcional subyacente a la respuesta bioquímica frente a este tipo de contaminantes. Resulta sorprendente que dicha respuesta sea prácticamente desconocida a nivel molecular, a pesar de su gran potencial aplicado en el contexto de la tecnología fitorremediadora. Para desarrollar este proyecto aplicamos a nuestros cultivos de chopo híbridos concentraciones diferentes de Aroclor 1221, una mezcla de PCBs muy utilizada a nivel comercial durante décadas, su uso está prohibido hoy internacionalmente. Y tomamos muestras de RNA a dos concentraciones y dos momentos distintos de exposición al contaminante, generando así una matriz de cuatro elementos con sus controles correspondientes. Con el fin de incrementar la especificidad de nuestro análisis, consideramos sobre todo los genes diferencialmente expresados más significativos según cuatro algoritmos estadísticos distintos. Por otra parte, realizamos análisis funcionales con herramientas bioinformáticas basadas en comparaciones de secuencias y en redes de co-expresión génica. La respuesta de los genes de particular interés fue validada mediante tecnología qRT-PCR (reacción de la polimerasa en cadena cuantitativa en tiempo real). Se trata del primer estudio comprehensivo de la respuesta de un organismo vegetal ante la presencia de PCBs. Este estudio nos ha permitido identificar una cantidad considerable de genes estructurales y reguladores, definiendo nuevos factores de transcripción cuya expresión es proporcional a la concentración de contaminante en el medio o al tiempo de exposición al mismo. Los análisis de correlación nos permiten afirmar en que la respuesta metabólica a PCBs, incluyendo posibles rutas degradadoras, participan en al menos quince factores de transcripción y unas cuarenta proteínas o enzimas que resultan particularmente inducidas. Entre las familias implicadas destacan los citocromos P450, la glutatión transferasas, las deshidrogenasas reductasas (short-chain dehydrogenase reductase) y las proteínas MDR (multi-drug resistance). Mientras que los factores de transcripción encontrados pertenecen a la familia de ZF-TF, MYBs, WRKYs entre otros. También identificamos proteínas de función desconocida que no se habían vinculado previamente a este tipo de respuestas en plantas, como la CSP (cold-shock domain proteins). Para estudiar su posible relación con la presencia de PCBs, se caracterizó un gen de esta familia detectado mediante espectrometría de masas en tándem (MS/MS) a partir de mapas IEF x SDS-PAGE (isoelectro focusing x sodium dodecyl sulphate- polyacrylamide gel electrophoresis) de alta resolución. Mediante qRT-PCR pudimos confirmar la inducción del gen correspondiente, ortólogo a PtCSP4 de P. trichocarpa (Potri.004g172600), en respuesta a Aroclor 1221. El análisis fenotípico de las líneas transgénicas de Arabidopsis thaliana que sobre-expresaba la proteína CSP de chopo híbrido confirmó un papel para la misma tolerancia a PCBs, posiblemente a través de mecanismos reguladores que activan proteínas MDR. Este trabajo, además de aportar datos novedosos sobre los mecanismos moleculares desencadenados por la presencia de un PCB en Populus, utilizado aquí como sistema modelo. Con ello se demuestra el potencial de las especies arbóreas no solo como agentes descontaminantes, ya explotado comercialmente, sino también como fuente potencial de genes interesantes. Entre los genes identificados en esta Tesis hay candidatos evidentes a participar en mecanismos de tolerancia al estrés inducido por la contaminación y también rutas metabólicas degradadores de PCBs. Precisamente la posibilidad de degradar al contaminante confiere particular interés a este tipo de estudios frente a la fitorremediación de metales pesados y otros contaminantes elementales. La comparación de los datos generados en este estudio con estudios análogos que se realicen en el futuro con otras especies y xenobióticos, contribuirán a definir mejor la respuesta de las plantas ante la contaminación orgánica y mejorar su potencial descontaminante. ABSTRACT Soil degradation has acquired a disturbing magnitude. Traditional methods of decontamination are expensive and insufficient. Phytoremediation represent an effective alternative, low cost, respectful of the environment, that also improves soil properties, although there have been relevant developments in the last decade. From a life scientist, the challenge is to decipher the major metabolic pathways involved in response to pollutants and understand their regulation. This information is essential if we desire to enhance the natural abilities of some plant species to remediate contaminated soils. This thesis studies have focused on Populus, the best available forestry model following the sequencing of the entire genome. Moreover, Populus has a natural ability to degrade organic pollutants, which explains its predominance in phytoremediation forestry programs currently being developed. We have chosen specifically to hybrid Populus tremula x P. alba, the ease with which it is grown and its particular biotechnological interest. This thesis presents a comprehensive study of the molecular response to polychlorinated biphenyls (PCBs), a family of persistent organic pollutants of particular relevance worldwide. It has been used for a transcriptomic approach using RNA-seq technology, to identify genes involved in the metabolism of compounds in plant and quantify their levels of activation in different controlled situations. The thesis also aims to define the underlying transcriptional control the biochemical response to these pollutants. It is surprising that the response is virtually unknown at the molecular level, despite its great potential applied in the context of phytoremediation technology. To develop this project we applied our hybrid poplar crops different concentrations of Aroclor 1221, a mixture of PCBs widely used commercially for decades, its use is now banned internationally. And we RNA samples at two different concentrations and times of exposure to the pollutant, generating an array of four elements with their corresponding controls. In order to increase the specificity of our analysis, we consider mainly the most significant differentially expressed genes in four different statistical algorithms. Moreover, functional analyzes conducted with bioinformatics tools based on sequence comparisons and networks gene co-expression. The response of genes of particular interest was validated by qRT-PCR (polymerase reaction chain in real-time quantitative. This is the first comprehensive study of the response of a plant organism in the presence of PCBs. This study allowed us to identify a considerable amount of structural and regulatory genes, defining new transcription factors whose expression is proportional to the concentration of contaminant in the middle or at the time of exposure. Correlation analyzes allow us to affirm that the metabolic response to PCBs, including possible degradative pathways, at least fifteen involved in transcription factors and forty proteins or enzymes which are particularly induced. Among the families involved include cytochromes P450, the glutathione transferases, dehydrogenases reductases (short -chain dehydrogenase reductase) and MDR proteins (multi - drug resistance). While transcription factors belong to the family found ZF-TF, MYBs, WRKYs among others. We also identify proteins of unknown function that had not been previously linked to such responses in plants such as CSP (cold- shock domain proteins). To study their possible relationship with the presence of PCBs, a gene in this family was characterized and was detected by tandem mass spectrometry (MS/MS) from maps IEF x SDS -PAGE (sodium dodecyl isoelectro x sulphate- polyacrylamide gel electrophoresis) of high resolution. By qRT -PCR could confirm the induction of the corresponding gene, ortholog to PtCSP4 of P. trichocarpa (Potri.004g172600), in response to Aroclor 1221. Phenotypic analysis of transgenic Arabidopsis thaliana lines over- expressing the protein CSP poplar hybrid confirmed a role for PCBs same tolerance, possibly through regulatory mechanisms activated MDR proteins. This work, in addition to providing new data on the molecular mechanisms triggered by the presence of PCBs in Populus, used here as a model system. Thus the potential of tree species not only as decontamination agents, and commercially exploited, but also as a potential source of interesting genes is shown. Among the genes identified in this thesis there are evident candidates to participate in tolerance mechanisms to stress induced by pollution and degrading metabolic pathways of PCBs. Precisely the possibility of degrading the pollutant attaches particular interest to this type of study off the phytoremediation of heavy metals and other elemental pollutants. The comparison of the data generated in this study with similar studies carried out in the future with other species and xenobiotics contribute to better define the response of plants to organic pollution and improve their decontamination potential.

Telemetría de vehículos FSAE con microcontrolador PIC32 y transceptor de radio CC1101

Quizás el campo de las telecomunicaciones sea uno de los campos en el que más se ha progresado en este último siglo y medio, con la ayuda de otros campos de la ciencia y la técnica tales como la computación, la física electrónica, y un gran número de disciplinas, que se han utilizado estos últimos 150 años en conjunción para mejorarse unas con la ayuda de otras. Por ejemplo, la química ayuda a comprender y mejorar campos como la medicina, que también a su vez se ve mejorada por los progresos en la electrónica creados por los físicos y químicos, que poseen herramientas más potentes para calcular y simular debido a los progresos computacionales. Otro de los campos que ha sufrido un gran avance en este último siglo es el de la automoción, aunque estancados en el motor de combustión, los vehículos han sufrido enormes cambios debido a la irrupción de los avances en la electrónica del automóvil con multitud de sistemas ya ampliamente integrados en los vehículos actuales. La Formula SAE® o Formula Student es una competición de diseño, organizada por la SAE International (Society of Automotive Engineers) para estudiantes de universidades de todo el mundo que promueve la ingeniería a través de una competición donde los miembros del equipo diseñan, construyen, desarrollan y compiten en un pequeño y potente monoplaza. En el ámbito educativo, evitando el sistema tradicional de clases magistrales, se introducen cambios en las metodologías de enseñanza y surge el proyecto de la Fórmula Student para lograr una mejora en las acciones formativas, que permitan ir incorporando nuevos objetivos y diseñar nuevas situaciones de aprendizaje que supongan una oportunidad para el desarrollo de competencias de los alumnos, mejorar su formación como ingenieros y contrastar sus progresos compitiendo con las mejores universidades del mundo. En este proyecto se pretende dotar a los alumnos de las escuelas de ingeniería de la UPM que desarrollan el vehículo de FSAE de una herramienta de telemetría con la que evaluar y probar comportamiento del vehículo de FSAE junto con sus subsistemas que ellos mismos diseñan, con el objetivo de evaluar el comportamiento, introducir mejoras, analizar resultados de una manera más rápida y cómoda, con el objetivo de poder progresar más rápidamente en su desarrollo, recibiendo y almacenando una realimentación directa e instantánea del funcionamiento mediante la lectura de los datos que circulan por el bus CAN del vehículo. También ofrece la posibilidad de inyectar datos a los sistemas conectados al bus CAN de manera remota. Se engloba en el conjunto de proyectos de la FSAE, más concretamente en los basados en la plataforma PIC32 y propone una solución conjunta con otros proyectos o también por sí sola. Para la ejecución del proyecto se fabricó una placa compuesta de dos placas de circuito impreso, la de la estación base que envía comandos, instrucciones y datos para inyectar en el bus CAN del vehículo mediante radiofrecuencia y la placa que incorpora el vehículo que envía las tramas que circulan por el bus CAN del vehículo con los identificadores deseados, ejecuta los comandos recibidos por radiofrecuencia y salva las tramas CAN en una memoria USB o SD Card. Las dos PCBs constituyen el hardware del proyecto. El software se compone de dos programas. Un programa para la PCB del vehículo que emite los datos a la estación base, codificado en lenguaje C con ayuda del entorno de desarrollo MPLAB de Microchip. El otro programa hecho con LabView para la PCB de la estación base que recibe los datos provenientes del vehículo y los interpreta. Se propone un hardware y una capa o funciones de software para los microcontroladores PIC32 (similar al de otros proyectos del FSAE) para la transmisión de las tramas del bus CAN del vehículo de manera inalámbrica a una estación base, capaz de insertar tramas en el bus CAN del vehículo enviadas desde la estación base. También almacena estas tramas CAN en un dispositivo USB o SD Card situado en el vehículo. Para la transmisión de los datos se hizo un estudio de las frecuencias de transmisión, la legislación aplicable y los tipos de transceptores. Se optó por utilizar la banda de radiofrecuencia de uso común ISM de 433MHz mediante el transceptor integrado CC110L de Texas Instruments altamente configurable y con interfaz SPI. Se adquirieron dos parejas de módulos compatibles, con amplificador de potencia o sin él. LabView controla la estación que recoge las tramas CAN vía RF y está dotada del mismo transceptor de radio junto con un puente de comunicaciones SPI-USB, al que se puede acceder de dos diferentes maneras, mediante librerías dll, o mediante NI-VISA con transferencias RAW-USB. La aplicación desarrollada posee una interfaz configurable por el usuario para la muestra de los futuros sensores o actuadores que se incorporen en el vehículo y es capaz de interpretar las tramas CAN, mostrarlas, gráfica, numéricamente y almacenar esta información, como si fuera el cuadro de instrumentos del vehículo. Existe una limitación de la velocidad global del sistema en forma de cuello de botella que se crea debido a las limitaciones del transceptor CC110L por lo que si no se desea filtrar los datos que se crean necesarios, sería necesario aumentar el número de canales de radio para altas ocupaciones del bus CAN. Debido a la pérdida de relaciones con el INSIA, no se pudo probar de manera real en el propio vehículo, pero se hicieron pruebas satisfactorias (hasta 1,6 km) con una configuración de tramas CAN estándar a una velocidad de transmisión de 1 Mbit/s y un tiempo de bit de 1 microsegundo. El periférico CAN del PIC32 se programará para cumplir con estas especificaciones de la ECU del vehículo, que se presupone que es la MS3 Sport de Bosch, de la que LabView interpretará las tramas CAN recibidas de manera inalámbrica. Para poder probar el sistema, ha sido necesario reutilizar el hardware y adaptar el software del primer prototipo creado, que emite tramas CAN preprogramadas con una latencia también programable y que simulará al bus CAN proporcionando los datos a transmitir por el sistema que incorpora el vehículo. Durante el desarrollo de este proyecto, en las etapas finales, el fabricante del puente de comunicaciones SPI-USB MCP2210 liberó una librería (dll) compatible y sin errores, por lo que se nos ofrecía una oportunidad interesante para la comparación de las velocidades de acceso al transceptor de radio, que se presuponía y se comprobó más eficiente que la solución ya hecha mediante NI-VISA. ABSTRACT. The Formula SAE competition is an international university applied to technological innovation in vehicles racing type formula, in which each team, made up of students, should design, construct and test a prototype each year within certain rules. The challenge of FSAE is that it is an educational project farther away than a master class. The goal of the present project is to make a tool for other students to use it in his projects related to FSAE to test and improve the vehicle, and, the improvements that can be provided by the electronics could be materialized in a victory and win the competition with this competitive advantage. A telemetry system was developed. It sends the data provided by the car’s CAN bus through a radio frequency transceiver and receive commands to execute on the system, it provides by a base station on the ground. Moreover, constant verification in real time of the status of the car or data parameters like the revolutions per minute, pressure from collectors, water temperature, and so on, can be accessed from the base station on the ground, so that, it could be possible to study the behaviour of the vehicle in early phases of the car development. A printed circuit board, composed of two boards, and two software programs in two different languages, have been developed, and built for the project implementation. The software utilized to design the PCB is Orcad10.5/Layout. The base station PCB on a PC receives data from the PCB connected to the vehicle’s CAN bus and sends commands like set CAN filters or masks, activate data logger or inject CAN frames. This PCB is connected to a PC via USB and contains a bridge USB-SPI to communicate with a similar transceiver on the vehicle PCB. LabView controls this part of the system. A special virtual Instrument (VI) had been created in order to add future new elements to the vehicle, is a dashboard, which reads the data passed from the main VI and represents them graphically to studying the behaviour of the car on track. In this special VI other alums can make modifications to accommodate the data provided from the vehicle CAN’s bus to new elements on the vehicle, show or save the CAN frames in the form or format they want. Two methods to access to SPI bus of CC110l RF transceiver over LabView have been developed with minimum changes between them. Access through NI-VISA (Virtual Instrument Software Architecture) which is a standard for configuring, programming, USB interfaces or other devices in National Instruments LabView. And access through DLL (dynamic link library) supplied by the manufacturer of the bridge USB-SPI, Microchip. Then the work is done in two forms, but the dll solution developed shows better behaviour, and increase the speed of the system because has less overload of the USB bus due to a better efficiency of the dll solution versus VISA solution. The PCB connected to the vehicle’s CAN bus receives commands from the base station PCB on a PC, and, acts in function of the command or execute actions like to inject packets into CAN bus or activate data logger. Also sends over RF the CAN frames present on the bus, which can be filtered, to avoid unnecessary radio emissions or overflowing the RF transceiver. This PCB consists of two basic pieces: A microcontroller with 32 bit architecture PIC32MX795F512L from Microchip and the radio transceiver integrated circuit CC110l from Texas Instruments. The PIC32MX795F512L has an integrated CAN and several peripherals like SPI controllers that are utilized to communicate with RF transceiver and SD Card. The USB controller on the PIC32 is utilized to store CAN data on a USB memory, and change notification peripheral is utilized like an external interrupt. Hardware for other peripherals is accessible. The software part of this PCB is coded in C with MPLAB from Microchip, and programming over PICkit 3 Programmer, also from Microchip. Some of his libraries have been modified to work properly with this project and other was created specifically for this project. In the phase for RF selection and design is made a study to clarify the general aspects of regulations for the this project in order to understand it and select the proper band, frequency, and radio transceiver for the activities developed in the project. From the different options available it selects a common use band ICM, with less regulation and free to emit with restrictions and disadvantages like high occupation. The transceiver utilized to transmit and receive the data CC110l is an integrated circuit which needs fewer components from Texas Instruments and it can be accessed through SPI bus. Basically is a state machine which changes his state whit commands received over an SPI bus or internal events. The transceiver has several programmable general purpose Inputs and outputs. These GPIOs are connected to PIC32 change notification input to generate an interrupt or connected to GPIO to MCP2210 USB-SPI bridge to inform to the base station for a packet received. A two pair of modules of CC110l radio module kit from different output power has been purchased which includes an antenna. This is to keep away from fabrication mistakes in RF hardware part or designs, although reference design and gerbers files are available on the webpage of the chip manufacturer. A neck bottle is present on the complete system, because the maximum data rate of CC110l transceiver is a half than CAN bus data rate, hence for high occupation of CAN bus is recommendable to filter the data or add more radio channels, because the buffers can’t sustain this load along the time. Unfortunately, during the development of the project, the relations with the INSIA, who develops the vehicle, was lost, for this reason, will be made impossible to test the final phases of the project like integration on the car, final test of integration, place of the antenna, enclosure of the electronics, connectors selection, etc. To test or evaluate the system, it was necessary to simulate the CAN bus with a hardware to feed the system with entry data. An early hardware prototype was adapted his software to send programed CAN frames at a fixed data rate and certain timing who simulate several levels of occupation of the CAN Bus. This CAN frames emulates the Bosch ECU MS3 Sport.

Análise dos parâmetros que interferem no metabolismo da microbiota anaeróbia e anóxica na remoção de bifenilas policloradas

A elevada concentração de cloro das bifenilas policloradas provoca alta toxicidade do composto, o qual dificulta sua biodegradação. A contaminação de PCB no Brasil foi confirmada em estudo realizado na Bahia de Santos-São Vicente (São Paulo), o qual revelou a necessidade de um plano de ação para o controle e remoção de PCB no Brasil. Pretendeu-se assim, na realização da presente pesquisa, verificar quatro hipóteses: (1) A técnica de Microextração em fase sólida é uma metodologia eficaz para avaliação de bifenilas policloradas de amostras de reatores; (2) A condição fermentativa-metanogênica abriga comunidade resistente ao PCB, e removê-lo; (3) A condição desnitrificante abriga comunidade resistente ao PCB, e removê-lo e (4) A remoção de PCB, bem como, a composição microbiana é distinta em cada condição metabólica. Para tanto, reatores em batelada foram montados separadamente com biomassa anaeróbia proveniente de reator UASB usado no tratamento de água residuária de avicultura e biomassa de sistemas de lodos ativados de tratamento de esgoto sanitário. Os reatores operados em condição mesófila foram alimentados com meio sintético, co-substratos, sendo etanol (457 mg.L-1) e formiato de sódio (680 mg.L-1) para os reatores anaeróbios, e somente etanol (598 mg.L-1) para os reatores anóxicos, além de PCB padrão Sigma (congêneres PCBs 10, 28, 52, 153, 138 e 180) em diferentes concentrações, dependendo do objetivo do ensaio. A aplicação do método de extração por SPME com análise em cromatógrafo gasoso com detector por captura de elétrons foi adequada para a determinação dos seis congêneres de PCB. Obteve-se ampla faixa de linearidade, seletividade frente aos vários interferentes, além da robustez do método, utilidade e confiabilidade na identificação e quantificação específica dos seis congêneres de PCB. A Hipótese 1 foi aceita; ou seja, por meio da aplicação da metodologia SPME foi possível quantificar os PCB nos reatores em batelada. Apesar de ter sido comprovada a inibição metanogênica na presença de PCB, com IC50 de 0,03 mg.LPCBs 10, 28, 52, 153, 138 e 180, respectivamente, para 1,5 mg.L-1 inicial. Thermotogaceae, Sedimentibacter, Anaerolinaceae e Pseudomonas, foram identificadas nos reatores anaeróbios por meio da plataforma Illumina. Representantes de Thermotogaceae e Sedimentibacter foram identificados em sistemas com elevada taxa de remoção de PCB, e representantes do filo Chloroflexi (grupo no qual representantes da Anaerolineae estão inseridos) foram os primeiros microrganismos desalogenadores de PCB identificados. Assim a Hipótese 2 foi aceita; ou seja, por meio de ensaios em batelada foi comprovada a toxicidade do PCB sobre a comunidade anaeróbia, a alteração da composição microbiana influenciada pela presença de PCB e a remoção nesta condição. Verificou-se ainda que na presença de PCB ocorreu a desnitrificação e comparando-se diferentes relações C/N-NO3-, foi estipulado a relação 6,95 como ideal na presença de PCB. Mesmo sendo confirmada a inibição da comunidade anóxica na presença de PCB com IC50 de 1,0 mg.L-1, verificou-se remoção de 1,02 mg.L-1, 0,85 mg.L-1, 1,31 mg.L-1, 1,02 mg.L-1, 0,03 mg.L-1, e 0,09 mg.L-1, para os PCBs 10, 28, 52, 153, 138 e 180, respectivamente, para 1,5 mg.L-1, inicial. Bactérias semelhantes a Aeromonadaceae, Lutispora, Sedimentibacter e Thermotogaceae foram identificadas nos reatores desnitrificantes. Representantes de Aeromonadaceae e Lutispora estão relacionados com o metabolismo desnitrificante e representantes de Thermotogaceae e Sedimentibacter foram identificados em sistemas com elevada taxa de remoção de PCB. A Hipótese 3 foi também aceita; ou seja, por meio de ensaios em batelada foi calculada a relação C/N-NO3- ideal na presença de PCB e foi comprovada a toxicidade do PCB sobre a comunidade anóxica, a alteração da composição microbiana influenciada pela presença de PCB e a remoção nesta condição. A maior remoção de PCB foi verificada para a condição anaeróbia (entre 45% a 100%), quando comparada com a condição anóxica (entre 10% a 95%). Bactérias semelhantes a Sedimentibacter e pertencentes a família Thermotogaceae foram identificadas nas duas condições nutricionais. Entretanto, mesmo verificando-se elevada abundancia relativa desses grupos nos reatores, evidenciou-se distinção entre as biomassas em cada condição. Assim, a Hipótese 4 foi aceita; ou seja, por meio de ensaios em batelada foi comprovada maior eficiência de remoção sob condição anaeróbia e distinta composição microbiana em cada condição.

Caracterização de valores de referência de qualidade de sedimentos para compostos orgânicos no sistema estuarino da baixada santista, São Paulo

Em regiões altamente contaminadas como a região da Baixada Santista, é importante estabelecer metas para a recuperação do ambiente. Apesar da ausência da contaminação ser a meta ideal, as implicações e os custos associados a esse objetivo, demanda o estabelecimento de metas de recuperação realistas em relação aos contaminantes presentes na região. Com o objetivo de caracterizar valores de referência de qualidade de sedimentos para compostos orgânicos na região da Baixada Santista, o Canal de Bertioga foi escolhido como local de referência por ser uma região sem fontes industrais ou outras fontes pontuais relativas aos compostos analisados. Amostras de água, sedimento e ostras foram coletadas e os hidrocarbonetos policíclicos aromáticos (PAHs), bifenilos policlorados (PCBs) e pesticidas organoclorados (OCPs) foram determinados por técnicas cromatográficas. A avaliação dos resultados de análises de PAHs, permite afirmar com alguma segurança, que os valores da somatória de PAHs das amostras sedimentos são, na sua grande maioria, inferiores a 1.000 µg/kg, não superando 1.600 µg/kg, concentrações abaixo dos limites estabelecidos na Resolução CONAMA 344/04 e abaixo dos valores que possam causar algum efeito adverso à biota, conforme valores descritos na literatura. Resultados de análise de PCBs, OCPs, compostos fenólicos e compostos orgânicos voláteis (VOCs) em amostras de sedimento, indicaram concentrações destes compostos abaixo dos limites de quantificação, exceto DDE (5,30 g/kg) e HCB (2,34 g/kg), que foram detectados em apenas um ii sítio de amostragem. Não houve evidências de possíveis fontes de emissão próximas à região de referência para PCBs, OCPs, compostos fenólicos e VOCs. Finalizando, espera-se que os resultados obtidos neste estudo possam fornecer subsídios para futuramente estabelecer uma área de referência para qualidade de sedimento na região da Baixada Santista, ou ainda serem utilizados em conjunto com as avaliações de contaminantes inorgânicos, testes ecotoxicológicos e indicadores biológicos, como ferramenta para avaliação da qualidade de sedimento e/ou para a classificação de material a ser dragado na região da Baixada Santista.

Reciclagem de resíduos eletroeletrônicos: um estudo cinético da lixiviação ácida de ferro de placas de circuito impresso de microcomputadores.

O aumento no consumo mundial de novos aparelhos eletroeletrônicos aliado à redução no tempo de vida útil destes equipamentos tem como principal consequência ao meio ambiente a geração de resíduos. No Brasil, com a instituição da Política Nacional de Resíduos Sólidos, criou-se a obrigatoriedade legal da responsabilidade dos fabricantes pela logística reversa dos equipamentos eletroeletrônicos, incentivando pesquisas para o desenvolvimento dos métodos de reciclagem e tratamento dos materiais descartados. O processo de lixiviação foi avaliado como alternativa à etapa de separação magnética presente nas atuais rotas hidrometalúrgicas para recuperação de metais valiosos de placas de circuito impresso. Para avaliar a composição das placas, foi realizado ensaio de dissolução em água régia. As amostras foram moídas e submetidas a ensaios de lixiviação com ácido sulfúrico nas concentrações de 1 e 2mol/L, às temperaturas de 75ºC, 85ºC e 95ºC, durante 24 horas. Com ácido sulfúrico 2mol/L a 95ºC, o tempo necessário para se obter 100% de extração do ferro foi de 2 horas. Nestas condições, não foi detectada a presença de cobre dissolvido. A cinética da reação é controlada por reação química e obedece a equação .=1(1)3. A energia de ativação aparente do processo equivale a 90kJ/mol.

Detección y análisis de contaminantes en diversos ambientes

Presentación realizada para Química Fusión, Valencia, 29 septiembre-1 octubre 2011.

Decomposition of two types of electric wires considering the effect of the metal in the production of pollutants

Combustion runs at 700 °C in a horizontal laboratory furnace were carried out on two different electric wires (PVC and halogen-free wire). Tests were performed in the presence and in the absence of the metal conductor of the wires. The analyses of the polycyclic aromatic hydrocarbons (PAHs), chlorobenzenes (CBzs), chlorophenols (CPhs), mono- to octa-chlorodibenzo-p-dioxin and dibenzofurans (PCDD/Fs), and dioxin-like PCBs are shown. Regarding semivolatile compounds, PAHs production decreases in the presence of metal, while a higher amount of chlorinated compounds are emitted. Respect to the PCDD/Fs, the PVC wire in the presence of metal presents the highest emission, with a much more emission of furans than dioxins. The maximum emission is with 2 or 3 chlorine atom PCDD/Fs. PCBs emission correlates with PCDD/F production and represents 3–4% of total toxicity, determined by using WHO2005 factors.

Pollutant emissions during pyrolysis and combustion of waste printed circuit boards, before and after metal removal

The constant increase in the production of electronic devices implies the need for an appropriate management of a growing number of waste electrical and electronic equipment. Thermal treatments represent an interesting alternative to recycle this kind of waste, but particular attention has to be paid to the potential emissions of toxic by-products. In this study, the emissions from thermal degradation of printed circuit boards (with and without metals) have been studied using a laboratory scale reactor, under oxidizing and inert atmosphere at 600 and 850 °C. Apart from carbon oxides, HBr was the main decomposition product, followed by high amounts of methane, ethylene, propylene, phenol and benzene. The maximum formation of PAHs was found in pyrolysis at 850 °C, naphthalene being the most abundant. High levels of 2-, 4-, 2,4-, 2,6- and 2,4,6-bromophenols were found, especially at 600 °C. Emissions of PCDD/Fs and dioxin-like PCBs were quite low and much lower than that of PBDD/Fs, due to the higher bromine content of the samples. Combustion at 600 °C was the run with the highest PBDD/F formation: the total content of eleven 2,3,7,8-substituted congeners (tetra- through heptaBDD/Fs) was 7240 and 3250 ng WHO2005-TEQ/kg sample, corresponding to the sample with and without metals, respectively.

Kinetic Analysis and Pollutant Evolution During Thermal Degradation of Printed Circuit Boards Considering the Effect of Metals

Paper submitted to the 7th International Symposium on Feedstock Recycling of Polymeric Materials (7th ISFR 2013), New Delhi, India, 23-26 October 2013.

Pollutant Formation During the Thermal Decomposition of Electrical and Electronic Wastes

Paper submitted to the 7th International Symposium on Feedstock Recycling of Polymeric Materials (7th ISFR 2013), New Delhi, India, 23-26 October 2013.

Analysis of pollutants evolved from combustion and pyrolysis of flexible polyurethane foam in a laboratory furnace

Resumen del póster presentado en PIC2015 – the 14th International Congress on Combustion By-Products and Their Health Effects, Umeå, Sweden, 14-17 June 2015.

Emissions of Gases, Polycyclic Aromatic Hydrocarbons and Brominated Pollutants during Thermal Degradation of Waste Printed Circuit Boards

Resumen del póster presentado en Symposium on Renewable Energy and Products from Biomass and Waste, CIUDEN (Cubillos de Sil, León, Spain), 12-13 May 2015

(Table 1) Neutral and phenolic organohalogen compounds in blubber samples of ringed seals (Pusa hispida) from East Greenland

We report on the comparative bioaccumulation, biotransformation and/or biomagnification from East Greenland ringed seal (Pusa hispida) blubber to polar bear (Ursus maritimus) tissues (adipose, liver and brain) of various classes and congeners of persistent chlorinated and brominated contaminants and metabolic by-products: polychlorinated biphenyls (PCBs), chlordanes (CHLs), hydroxyl (OH-) and methylsulfonyl (MeSO2-) PCBs, polybrominated biphenyls (PBBs), OH-PBBs, polybrominated diphenyl ether (PBDE) and hexabromocyclododecane (HBCD) flame retardants and OH- and methoxyl (MeO-) PBDEs, 2,2-dichloro-bis(4-chlorophenyl)ethene (p,p'-DDE), 3-MeSO2-p,p'-DDE, pentachlorophenol (PCP) and 4-OH-heptachlorostyrene (4-OH-HpCS). We detected all of the investigated contaminants in ringed seal blubber with high frequency, the main diet of East Greenland bears, with the exception of OH-PCBs and 4-OH-HpCS, which indicated that these phenolic contaminants were likely of metabolic origin and formed in the bears from accumulated PCBs and octachlorostyrene (OCS), respectively, rather than being bioaccumulated from a seal blubber diet. For all of the detectable sum of classes or individual organohalogens, in general, the ringed seal to polar bear mean BMFs for SumPCBs, p,p'-DDE, SumCHLs, SumMeSO2-PCBs, 3-MeSO2-p,p'-DDE, PCP, SumPBDEs, total-(alpha)-HBCD, SumOH-PBDEs, SumMeO-PBDEs and SumOH-PBBs indicated that these organohalogens bioaccumulate, and in some cases there was tissue-specific biomagnification, e.g., BMFs for bear adipose and liver ranged from 2 to 570. The blood-brain barrier appeared to be effective in minimizing brain accumulation as BMFs were <= 1 in the brain, with the exception of SumOH-PBBs (mean BMF = 93±54). Unlike OH-PCB metabolites, OH-PBDEs in the bear tissues appeared to be mainly accumulated from the seal blubber rather than being metabolic formed from PBDEs in the bears. In vitro PBDE depletion assays using polar bear hepatic microsomes, wherein the rate of oxidative metabolism of PBDE congeners was very slow, supported the probability that accumulation from seals is the main source of OH-PBDEs in the bear tissues. Our findings demonstrated from ringed seal to polar bears that organohalogen biotransformation, bioaccumulation and/or biomagnification varied widely and depended on the contaminant in question. Our results show the increasing complexity of bioaccumulated and in some cases biomagnified, chlorinated and brominated contaminants and/or metabolites from the diet may be a contributing stress factor in the health of East Greenland polar bears.

Thyroid hormones and halogenated organic compounds in black-legged kittiwake (Rissa tridactyla) and northern fulmar (Fulmarus glacialis)

Thyroid hormones are essential for normal growth and development and disruption of thyroid homeostasis can be critical to young developing individuals. The aim of the present study was to assess plasma concentrations of halogenated organic contaminants (HOCs) in chicks of two seabird species and to investigate possible correlations of HOCs with circulating thyroid hormone (TH) concentrations. Plasma from black-legged kittiwake (Rissa tridactyla) and northern fulmar (Fulmarus glacialis) chicks were sampled in Kongsfjorden, Svalbard in 2006. The samples were analyzed for thyroid hormones and a wide range of HOCs (polychlorinated biphenyls (PCBs), hydroxylated (OH-) and methylsulphoned (MeSO-) PCB metabolites, organochlorine pesticides (OCPs), brominated flame retardants (BFRs), and perfluorinated compounds (PFCs)). Concentrations of HOCs were generally low in kittiwake and fulmar chicks compared to previous reports. HOC concentrations were five times higher in fulmar chicks compared to in kittiwake chicks. PFCs dominated the summed HOCs concentrations in both species (77% in kittiwakes and 69% in fulmars). Positive associations between total thyroxin (TT4) and PFCs (PFHpS, PFOS, PFNA) were found in both species. Although correlations do not implicate causal relationships per se, the correlations are of concern as disruption of TH homeostasis may cause developmental effects in young birds.

(Table 1) Organohalogen contaminants in adipose tissue of captive sledge dogs (Canis familiaris) fed with whale blubber or pork fat

Studies on the fate of organohalogen contaminants (OHCs) in wild top predator mammals in the Arctic have often been a challenge due to important knowledge deficiencies in the life history of the sampled animals. The present study investigated the influence of age, dietary and trans-generational factors on the fate of major lipophilic chlorinated and brominated OHCs in adipose tissue of a potential surrogate captive species for the polar bear (Ursus maritimus), the sledge dog (Canis familiaris) in West Greenland. Adult female sledge dogs (P) and their sexually-mature (F1) and/or pre-weaning pups (F1-MLK) were divided into an exposed group (EXP) fed blubber from a Greenland minke whale (Balaenoptera acutorostrata) and a control group (CON) given commercially available pork fat. Large dietary treatment-related differences in summed and individual congener/compound adipose tissue concentrations of polybrominated diphenyl ethers (PBDEs), hexachlorobenzene (HCB), chlordanes (CHLs) and polychlorinated biphenyls (PCBs) were found between the EXP and CON groups for all the sledge dog cohorts. However, among the F1-MLK, F1 and P dogs in both of the EXP and CON groups, little or no difference existed in PBDE, HCB, CHL and PCB concentrations, suggesting higher state of equilibrium in adipose tissue concentrations from a very early stage of life. In contrast, the distribution pattern (proportions to the summed concentrations) of OHC classes, and the major congeners/ compounds constituting those classes, varied on a dietary group- and/or cohort-dependent manner. The present captive sledge dog study demonstrated the importance of the confounding effects of diet composition, mother-pup association (maternal transfer), reproductive status (nursing), and to a lesser extent age in the fate of OHCs in adipose tissue of a large top carnivore mammal.