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Élaboration d'une approche de biosurveillance humaine pour évaluer l'exposition aux métaux et éléments traces de la population libanaise et son association avec la consommation d'eau potableNasser Eddine, Nessrine 08 1900 (has links)
Dans le contexte libanais, les infrastructures de traitement et d’approvisionnement en eau potable ne sont pas optimales et il existe un manque de connaissances sur l’association entre la consommation d’eau et les niveaux d’exposition aux métaux et éléments traces. La présente étude est une première tentative faisant état de l'exposition aux métaux et aux éléments traces dans des sous-groupes de la population libanaise en utilisant une approche de biosurveillance multi-matrices. Les concentrations en 11 métaux et éléments traces (aluminium (Al), arsenic (As), cadmium (Cd), chrome (Cr), cuivre (Cu), fer (Fe), plomb (Pb), manganèse (Mn), sélénium (Se), uranium (U), zinc (Zn)) ont été mesurées dans des échantillons d'urine, de cheveux et d'ongles d’orteils d'un groupe de la population et les niveaux ont été comparés en fonction de l'âge, du sexe, du tabagisme, du statut économique, de la zone géographique et de la source d'eau potable. Alors que la plupart des concentrations urinaires et d'ongles n'étaient pas statistiquement différentes entre les hommes et les femmes, les concentrations mesurées d'éléments dans les cheveux étaient statistiquement plus élevées chez les femmes que chez les hommes. Les concentrations urinaires d'Al, Cu, Se et Zn étaient statistiquement plus élevées chez les enfants comparativement aux adolescents et aux adultes. Les concentrations de plusieurs éléments dans les cheveux et les ongles (As, Cd, Pb, Mn, Se dans les cheveux et les ongles plus Al, Fe dans les ongles d’orteils) étaient significativement plus élevées chez les enfants que chez les adolescents et/ou adultes (MG chez les enfants vs. les adolescents vs. les adultes dans les cheveux: 0,009 vs 0,009 vs. 0.005 µg As/g (p<0,01); 0,046 vs. 0,016 vs. 0,022 µg Cd/ g (p<0,05); 1,16 vs. 0,580 vs. 0,627 µg Pb/g (p<0,1); 0,305 vs. 0,168 vs. 0,275 µg Mn/g (p<0,1); 0,365 vs. 0,370 vs. 0,217 µg Se/g (p<0,05) et dans les ongles d’orteils : 17,0 vs. 14,3 vs. 7,31 µg Al/g et 56,9 vs. 46,0 vs. 23,8 µg Fe/g (p<0,05). Le statut tabagique n’a aucune influence sur les concentrations de As, cd et Pb. Les niveaux de Cd, Pb et Mn étaient également statistiquement plus élevés dans les échantillons des sous-groupes ayant un statut économique inférieur (Cd et Pb dans les 3 matrices urines, cheveux et ongles d’orteils respectivement de 0,223 µg Cd/L(p<0,1); 0,062 µg Cd/ g (p<0,05) et 0,038 µg Cd/ g (p<0,01); 1,16 µg Pb/L(p<0,05); 1,84 µg Pb/ g (p<0,01) et 0,840 µg Pb/ g (p<0,01) et Mn dans les cheveux et ongles d’orteils respectivement de 0,434 µg Mn/ g (p<0,05); 0,689 µg Mn/g (p<0,05). Les mêmes tendances ont été observées pour la stratification en fonction de la zone géographique, mais la différence n'était pas statistiquement significative, sauf pour le Pb dans l'urine où les participants de Beyrouth centre avaient des niveaux urinaires significativement moins élevés que les autres zones (0,560 µg Pb/L(p<0,01). Très peu de corrélations ont été identifiées entre les sources d’eau potable et les concentrations des métaux et éléments traces dans les urines, cheveux et ongles d’orteils. Cependant, une corrélation a été trouvée entre les niveaux d’As, Cd et Pb dans les cheveux et les ongles d’orteils respectivement (r = 0,4, p<0,05); (r = 0,310, p<0,05) et (r = 0.,270, p<0,1). Dans l’ensemble, les résultats ont montré que la population était exposée à des concentrations plus élevées de certains métaux que les autres populations, une attention particulière doit être accordée à l'exposition aux As, Cd et Pb, Mn et Se. Bien que la source d'eau n'ait pas contribué à la différence dans les niveaux d'exposition, les niveaux de certains métaux et éléments traces différaient selon l'âge, le sexe, les zones géographiques de résidence et le statut économique. Il pourrait être pertinent d'étendre ce type d'enquête à une initiative de biosurveillance humaine à grande échelle dans la population libanaise afin de valider et de généraliser les résultats et d'observer les tendances temporelles dans le temps. / In the Lebanese context, drinking water treatment and supply infrastructures are not optimal and
it exists a lack of knowledge on the association between water consumption and levels of exposure
to metals and trace elements. The present study is a first attempt reporting exposure to metals and
trace elements in subgroups of the Lebanese population using a multi-matrix biomonitoring
approach. Concentrations of 11 metals and trace elements (aluminum (Al), arsenic (As), cadmium
(Cd), chromium (Cr), copper (Cu), iron (Fe), lead (Pb), manganese (Mn), selenium (Se), uranium
(U), zinc (Zn)) were measured in urine, hair and toenail samples from a population group and the
levels were compared according to the age, gender, smoking status, economic status, geographic
area, and source of drinking water. While most urinary and nail concentrations were not
statistically different between men and women, measured element concentrations in hair were
statistically higher in women than in men. Urinary concentrations of Al, Cu, Se and Zn were
statistically higher in children compared to adolescents and adults. Concentrations of several
elements in hair and nails (As, Cd, Pb, Mn, Se in hair and toenails plus Al, Fe in toenails) were
significantly higher in children than in adolescents and/or adults (MG in children vs. adolescents
vs. adults in hair: 0.009 vs. 0.009 vs. 0.005 µg As/g (p<0.01); 0.046 vs. 0.016 vs. 0.022 µg Cd/g
(p< 0.05), 1.16 vs. 0.580 vs. 0.627 µg Pb/g (p<0.1), 0.305 vs. 0.168 vs. 0.275 µg Mn/g (p<0.1),
0.365 vs. 0.370 vs. 0.217 µg Se/g (p<0.05) and in the nails: 17.0 vs. 14.3 vs. 7.31 µg Al/g and 56.9
vs. 46.0 vs. 23.8 µg Fe/g. Smoking status has no influence on the concentrations of metals and
trace elements. The levels of Cd, Pb and Mn were also statistically higher in the samples of the
subgroups with a lower economic status (Cd and Pb in the 3 matrices urine, hair and toenails
respectively of 0.223 µg Cd/L (p<0, 1);0.062 µg Cd/g (p<0.05) and 0.038 µg Cd/g (p<0.01);1.16
µg Pb/L(p<0.05);1.84 µg Pb/ g (p<0.01) and 0.840 µg Pb/g (p<0.01) and Mn in hair and toenails
respectively 0.434 µg Mn/g (p<0.05), 0.689 µg Mn/g (p <0.05). The same trends were observed
for stratification according to geographical area, but the difference was not statistically significant,
except for Pb in urine where participants from central Beirut had significantly lower urinary levels
than the others zones (0.560 µg Pb/L (p<0.01). Very few correlations have been identified between
the sources of drinking water and the concentrations of metals and trace elements in urine, hair
and toenails. However, a correlation was found between As, Cd and Pb levels in hair and toenails
respectively r = 0.4, p<0.05), r = 0.310, p<0.05) and r = 0.270, p<0.1). Overall, the results showed
that the population was exposed to higher concentrations of some metals than other populations, particular attention should be paid to exposure to As, Cd and Pb, Mn and Se. Although water
source did not contribute to the difference in exposure levels, levels of some metals and trace
elements differed by age, sex, smoking status, geographic areas of residence, and economic status.
It might be relevant to extend this type of investigation to a large-scale human biomonitoring
initiative in the Lebanese population in order to validate and generalize the results and to observe
temporal trends over time.
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A Computer-Based Decision Tool for Prioritizing the Reduction of Airborne Chemical Emissions from Canadian Oil Refineries Using Estimated Health ImpactsGower, Stephanie Karen January 2007 (has links)
Petroleum refineries emit a variety of airborne substances which may be harmful to human health. HEIDI II (Health Effects Indicators Decision Index II) is a computer-based decision analysis tool which assesses airborne emissions from Canada's oil refineries for reduction, based on ordinal ranking of estimated health impacts. The model was designed by a project team within NERAM (Network for Environmental Risk Assessment and Management) and assembled with significant stakeholder consultation. HEIDI II is publicly available as a deterministic Excel-based tool which ranks 31 air pollutants based on predicted disease incidence or estimated DALYS (disability adjusted life years). The model includes calculations to account for average annual emissions, ambient concentrations, stack height, meteorology/dispersion, photodegradation, and the population distribution around each refinery. Different formulations of continuous dose-response functions were applied to nonthreshold-acting air toxics, threshold-acting air toxics, and nonthreshold-acting CACs (criteria air contaminants). An updated probabilistic version of HEIDI II was developed using Matlab code to account for parameter uncertainty and identify key leverage variables. Sensitivity analyses indicate that parameter uncertainty in the model variables for annual emissions and for concentration-response/toxicological slopes have the greatest leverage on predicted health impacts. Scenario analyses suggest that the geographic distribution of population density around a refinery site is an important predictor of total health impact. Several ranking metrics (predicted case incidence, simple DALY, and complex DALY) and ordinal ranking approaches (deterministic model, average from Monte Carlo simulation, test of stochastic dominance) were used to identify priority substances for reduction; the results were similar in each case. The predicted impacts of primary and secondary particulate matter (PM) consistently outweighed those of the air toxics. Nickel, PAH (polycyclic aromatic hydrocarbons), BTEX (benzene, toluene, ethylbenzene and xylene), sulphuric acid, and vanadium were consistently identified as priority air toxics at refineries where they were reported emissions. For many substances, the difference in rank order is indeterminate when parametric uncertainty and variability are considered.
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The Distribution of Environmental Contaminants: a Socio-Historical Study of Selected Neighborhoods in Dallas County, TexasCutrer, Jennifer G. 12 1900 (has links)
This research expands on recent sociological studies which maintain that environmental contaminants in America are disproportionately placed in neighborhoods inhabited by minorities and the poor. Prior studies have focused on the predictor variables which identify areas of contamination near residential neighborhoods, yet fail to explore the socio-political and historical factors which contribute to these phenomena. The Environmental Protection Agency's Toxic Release Inventory 1990 database, the Texas Natural Resource Conservation Commission's Annual Report of the Hazardous and Solid Waste Program for 1992, and the U.S. Census Bureau's 1990 Census Data for Dallas County were utilized in pinpointing industries violating toxic release standards. Socio-historical data was obtained from government and historical records and reports, books, and newspaper clippings on Dallas County. Maps and data were obtained from the North Central Texas Council of Governments, and the cities of Dallas and Garland. Chapter I discusses the synergetic forces of capitalism, urban growth, uneven development, and settlement patterns resulting in the distribution of environmental contaminants. Chapter II reviews the literature and presents evidence that race and class are strong predictors of where environmental contaminants are located. Chapter III outlines the data and methods employed. Chapter IV traces the historical development of Dallas County. Chapter V details those political, economic, and social factors contributing to the convergence of people and contaminants within three selected neighborhoods. The forces which historically relegate minorities, particularly Blacks and Hispanics, and the poor to less desirable jobs, cheaper housing, and land costs are also explored. Cheap land and labor attract industry which, in turn, attracts more laborers. Chapter VI, the summary and conclusions, utilizes the socio-spatial approach in examining urban infrastructure development (i.e. roads and railways) which also reduces adjacent land costs making housing more affordable for minorities and the poor. This study concludes that because of historical development and capitalism's exploitation of labor, these populations become entrenched in neighborhoods located adjacent to those industries where contaminants are emitted.
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Biochemical and molecular characterization of heavy metal resistant bacteria isolated from the Klip River, South AfricaChihomvu, Patience January 2014 (has links)
M. Tech. (Department of Biotechnology, Faculty of Engineering and Technology) Vaal University of Technology / The Klip River has suffered severe anthropogenic effects from industrial, agricultural, mining and domestic activities. As a result harmful contaminants such as heavy metals have accumulated in the river, causing microorganisms inhabiting the environment to develop mechanisms to protect them from the harmful effects of the contaminants. The current study deals with the isolation and characterization of heavy metal resistant bacteria isolated from the Klip River Catchment. Water and sediment samples were collected from 6 sites of the Klip River, and the Vaal Barrage (control). In-situ parameters, such as pH, turbidity, salinity, conductivity, temperature and dissolved oxygen were determined. Lead, iron, cadmium, nickel, zinc and copper concentrations of water were determined by atomic absorption spectroscopy. For bacterial analysis sediment and water samples were collected in sterile glass jars and bottles respectively. Heavy metal resistant bacterial isolates were screened on heavy metal constituted Luria Bertani (LB) agar. Biochemical profiles of the isolates were constructed using the API 20E® strips, antibiotic susceptibility tests were done and growth studies were carried out using spectrophotometric methods. The isolates were identified using 16SrDNA sequencing and alignment.
A partial sequence of the copper resistance gene pcoA was amplified from strains Lysinibacillus sp. KR25 [KJ935917], and Escherichia coli KR29 [KJ935918]. The pcoR gene was amplified from E. coli (KR29) and the partial sequence for the chromate resistance gene chrB, was amplified from Pseudomonas sp. KR23 [KJ935916]. The gene fragments were then sequenced and translated into protein sequences. The partial protein sequences were aligned with existing copper and chromate resistance proteins in the Genbank and phylogenetic analysis was carried out. The physico-chemical properties of the translated proteins were predicted using the bioinformatics tool Expasy ProtParam Program. A homology modelling method was used for the prediction of secondary structures using SOPMA software, 3D-protein modelling was carried out using I-TASSER. Validation of the 3D structures produced was performed using Ramachandran plot analysis using MolProbity, C-score and TM-scores. Plasmid isolation was also carried out for both the wild type strains and cured derivatives and their plasmid profiles were analysed using gel electrophoresis to ascertain the presence of plasmids in the isolates. The cured derivatives were also plated on heavy metal constituted media. Antibiotic disc diffusion tests were also carried out to ascertain whether the antibiotic resistance determinants were present on the plasmid or the chromosome.
The uppermost part of the Klip River had the lowest pH and thus the highest levels of heavy metal concentrations were recorded in the water samples. Turbidity, salinity and specific conductivity increased measurably at Site 4 (Henley on Klip Weir). Sixteen isolates exhibiting high iron and lead resistance (4 mM) were selected for further studies. Antibiotic susceptibility tests revealed that the isolates exhibited multi-tolerances to drugs such as Ampicillin (10 μg/ml), Amoxcyllin (10 μg/ml), Cephalothin acid (30 μg/ml), Cotrimoxazole (25 μg/ml), Neomycin (30 μg/ml), Streptomycin (10 μg/ml), Tetracycline (30 μg/ml), Tobramycin (10 μg/ml) and Vancomycin (30 μg/ml). Growth studies illustrated the effect of heavy metals on the isolates growth patterns. Cadmium and chromium inhibited the growth of most of the microorganisms. The following strains had high mean specific growth rates; KR01, KR17, and KR25, therefore these isolates have great potential for bioremediative applications.
Using 16SrDNA sequencing the isolates were identified as KR01 (Aeromonas hydrophila), KR02 (Bacillus sp.), KR04 (Bacillus megaterium), KR06 (Bacillus subtilis), KR07 (Pseudomonas sp), KR17 (Proteus penneri), KR18 (Shewanella), KR19 (Aeromonas sp.), KR22 (Proteus sp.), KR23 (Pseudomonas sp.), KR25 (Lysinibacillus sp.), KR29 (Escherichia coli), KR44 (Bacillus licheniformis) and KR48 (Arthrobacter sp.).
Three heavy metal resistance genes were detected from three isolates. The pcoA gene was amplified from strains Lysinibacillus sp KR25, and Escherichia coli KR29; pcoR gene from E. coli KR29 and the chrB gene, from Pseudomonas sp. KR23. The genes encoding for heavy metal resistance and antibiotic resistance were found to be located on the chromosome for both Pseudomonas sp. (KR23) and E.coli (KR29). For Lysinibacillus (KR25) the heavy metal resistance determinants are suspected to be located on a mobile genetic element which was not detected using gel electrophoresis. The translated protein sequence for pcoA_25 showed 82% homology with the copper resistant protein form Cronobacter turicensis [YP003212800.1]. Sequence comparisons between the pcoR partial protein sequence found in E. coli KR29 showed 100% homology with 36 amino acids (which was 20% of the query cover) from a transcriptional regulatory protein pcoR found in E. coli [WP014641166.1]. For the chrB partial protein sequence detected in Pseudomonas sp. (KR23), 97% of the query sequence showed 99% homology to a vitamin B12 transporter btuB in Stenotrophus sp. RIT309.
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Desenvolvimento de um método para determinação de disruptores endócrinos em saliva por microextração líquido-líquido dispersiva e cromatografia líquida de alta eficiência acoplada à espectrometria de massas / Method development for determination of endocrine disrupting in saliva by dispersive liquid-liquid microextraction and high performance liquid chromatography-tandem mass spectrometryOliveira, Mariana Lepri de 11 May 2018 (has links)
A crescente demanda por novos compostos químicos sintéticos para atender as necessidades médicas, agrícolas e industriais da população resultou no surgimento de novos contaminantes, denominados emergentes. Com a intensa utilização destes compostos em diversos produtos e o consequente aumento da concentração destes no meio ambiente, a população passou a estar exposta e sujeita aos seus possíveis efeitos deletérios. Os disruptores endócrinos fazem parte do grupo de contaminantes emergentes que tem a capacidade de interferir na ação natural dos hormônios por diferentes mecanismos, podendo agravar o quadro de doenças pré-existentes e causar diversos outros danos à saúde humana. Apesar de serem largamente utilizados na vida cotidiana, a regulamentação do uso desses compostos ainda é quase inexistente, aumentando a preocupação quanto à sua segurança. Comumente, os estudos de biomonitoramento humano para avaliar a exposição da população em geral são realizados utilizando amostras de sangue e urina. Neste sentido, o presente estudo teve como objetivo desenvolver um método alternativo para determinação simultânea de 17 compostos classificados como disruptores endócrinos: bisfenóis, parabenos, benzofenonas e triclocarban em saliva, empregando a microextração líquido-liquido dispersiva (DLLME) no preparo da amostra e cromatografia líquida de alta eficiência acoplada à espectrometria de massas (LC-MS/MS) para a determinação. Os seguintes parâmetros foram avaliados na otimização da DLLME: volume e tipo de solvente extrator e dispersor, força iônica, pH e agitação. Após a otimização, o processo de extração foi realizado adicionando-se 500 ?L de água deionizada à 500 ?L da amostra de saliva. Os solventes extrator e dispersor selecionados foram triclorometano e acetona, respectivamente (volume de 2 mL da mistura na proporção 1:3). Foi adicionada a etapa de agitação manual por dez segundos. As curvas de calibração de todos os analitos apresentaram linearidade, com coeficiente de correlação superiores a 0,99 no intervalo de concentração de 1-20 ng mL-1. O coeficiente de variação (CV) intra e interdias foram inferiores a 20% para o primeiro ponto da faixa linear e inferiores a 15% para as demais concentrações. O erro padrão relativo intra e interdias foram inferiores a 20% para o primeiro ponto da faixa linear e inferiores a 15% nas demais concentrações. Os limites de detecção (LOD) e de quantificação (LOQ) encontrados variaram de 0,01 - 0,15 ng mL-1 e 0,05 - 0,40 ng mL-1, respectivamente. Para avaliar a aplicabilidade do método, dez amostras de saliva foram analisadas e apresentaram concentrações detectáveis de bisfenol S (BPS), bisfenol AF (BPAF) e propilparabeno (PrP) ( <LOQ). O bisfenol A (BPA) e a benzofenona-3 (BP3) foram encontrados em concentrações que variaram entre 1,01-3,14 e 1,00-2,50 ng mL-1, respectivamente. O metilparabeno (MeP) foi detectado em todas as amostras, atingindo, nos níveis quantificáveis, a concentração de 17,8 ng mL-1. Portanto, o método aqui proposto é uma alternativa simples e rápida para estudos de biomonitoramento humano, capaz de determinar 17 analitos multi-classes simultaneamente com uso de baixo volume de solventes. / The growing demand of new synthetic chemical compounds to attend the medical, agricultural and industrial needs of the population has resulted in the emergence of new contaminants, known as emerging contaminants. The presence of these contaminants in several products, and its consequent concentration increase in the environment, exposed the population and subject them to possible deleterious effects. Endocrine disruptors are a group of these contaminants that have the ability to interfere at the natural action of hormones by different mechanisms, wich can worsen preexistent diseases and cause other damages to humam health. Despite the widely utilization of these compounds in everyday life, regulation of its use is almost non-existent, raising concerns about their safety. Mostly human biomonitoring studies to assess the exposure of the population are performed in urine and blood samples. In these sense, the present study aimed to develop an alternative method for simultaneous determination of 17 endocrine disrupting compounds: bisphenols, parabens, benzophenones and triclocarban in saliva, using dispersive liquid-liquid microextraction (DLLME) at sample preparation and high performance liquid chromatography coupled to mass spectrometry (LC-MS/MS) for determination. The evaluated parameters in the optimization of DLLME were: volume and type of extraction and disperser solvents, ionic strength, pH and agitation. After the optimization, the extraction procedure was performed by adding 500 ?L of deionized water to 500 ?L of saliva sample. The extraction and disperser solvents selected were trichloromethane and acetone, respectively (2 mL of the mixture in 1:3 ratio). The manual agitation for ten seconds. The matrix-matched calibration curves of all analytes were linear, with correlation coefficients higher than 0.99 in the range level of 1 - 20 ng mL-1. The intra and interday coefficient of variation (CV) was less than 20% for the first point of the linear range and less than 15% for the other concentrations. The intra and interday relative standard deviation (RSD) was less than 20% for the first point of the linear range and less than 15% for the other concentrations. The limits of detection (LOD) and quantification (LOQ) ranged from 0,01 - 0,15 ng mL-1 and 0,05 - 0,4 ng mL-1, respectively. To evaluate the applicability of the method, ten saliva samples were analyzed, showing detectable concentrations (<LOQ) of bisphenol S (BPS), bisphenol AF (BPAF) and propylparaben (PrP). Bisphenol A (BPA) and benzophenone-3 (BP3) were found in ranging concentrations from 1,01 - 3,14 and 1,00 - 2,50 ng mL-1, respectively. Methylparaben (MeP) was detected in all samples, reaching, at the quantifiable levels, the concentration of 17,8 ng mL-1. Thus, the proposed method is a simple and fast alternative for human biomonitoring studies, capable of determining 17 multi-class analytes simultaneously using low volume of solvent.
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A study of the performance of biochar as adsorbing agent in o‐DGT devices / En studie av egenskaperna hos biokol som adsorberande agent i o‐DGT anordningarEliasson, Anna January 2017 (has links)
A new complex aspect in the matter of water quality is the occurrence of emerging organic pollutants and contaminants in waste water. The currently low extent to which treatment of waste water is performed in Brazil, and in the world as a whole, there is a considerable need for development of cheap and accurate in-situ sampling methods for far-reaching studies of surface water quality. The lack of such methods today makes the maintenance and establishing of sanitary safety difficult. This diploma work gives a brief introduction to the basic principles of the passive sampling method known as Diffusive Gradient in Thin-films (DGT). A method that could be useful for such monitoring of quality in water bodies world wide. The aim of this study is to develop a method, for the detection of organic emerging pollutants and contaminants – i.e. compounds, which usually are present at very low concentrations when found in the environment as a result of human activity. More specifically, this work investigates the potential and usefulness of the application of DGT devices in detection of organic compounds that can affect human health and ecosystems, even at low concentrations, however, their effects still are in need of further investigations. This study focuses on both purely technical as well as practical points of views. The efficiency of organic DGT (o-DGT) with biochar as the adsorbing agent is examined targeting the detection of organic pollutants and contaminants in surface water. In this sense, the specific aim of the work is to evaluate the performance of biochar as adsorbing agent. This work showed that the performance of biochar as the adsorbing agent in binding layers in o-DGT sample devices can be considered as satisfactory since all compounds of interest in this study was successfully detected, quantified an identified. Further investigations in the future are needed to determine the effects of varying pH, temperature and ion concentration in the deployment media, as well as the properties of the binding layer in relation to concentration of biochar and the thickness of the layer. These in order to optimize the method for in-situ water sampling, aiming conventional use of biochar as the adsorbing agent in the future.
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Monitoramento temporal e espacial de contaminações bacterianas na produção de bioetanol: caracterização molecular por T-RFLP e detecção quantitativa por qPCRde comunidades formadoras de biofilmes / Temporal and spatial monitoring of bacterial contamination in bioethanol production: a molecular characterization by T-RFLP and quantitative detection by qPCR of community-formers biofilmsCampana, Felippe Buck 14 September 2012 (has links)
A contaminação bacteriana por espécies dos gêneros Lactobacillus, Bacillus e Leuconostoc entre outras bactérias lácticas é um dos principais fatores que afetam o rendimento da fermentação alcoólica. A formação de biofilmes acaba protegendo as bactérias e é uma fonte permanente de contaminação. Objetivando caracterizar tais contaminações em (1) biofilmes de centrífuga, dorna, trocador de calor e tubulação de água e (2) melaço, mosto, levedo, levedo tratado (com H2SO4) e vinho, amostras foram coletadas em diferentes períodos de um sistema fermentativo de alto teor alcoólico (16%). As enzimas de restrição AluI, BstUI, HaeIII, HinfI, MseI e MspI utilizadas nas análises de T-RFLP foram definidas por análises in silico com sequências do gene 16S rRNA de contaminantes freqüentes. Essas enzimas geram uma maior quantidade de T-RFs únicos entre 30 e 650 pb. Os DNAs extraídos das amostras foram submetidos às análises de T-RFLP para obtenção do perfil molecular das comunidades microbianas dos pontos de coleta. Os índices de diversidade de Shannon foram calculados com base no número dos T-RFs. Foram realizadas as análises dos componentes principais (PCA) e a inferência filogenética dos contaminantes com base nos perfis dos T-RFs. A quantificação dos principais táxons contaminantes foi feita por qPCR utilizando primers específicos delineados neste estudo e considerando a média de cópias do gene 16S rRNA presentes no genoma de cada táxon bacteriano. Na primeira coleta o biofilme de água apresentou maior índice de diversidade microbiana e na segunda, melaço e mosto. PCA sugere que os biofilmes (e não as fontes externas) são os principais contaminantes desse processo fermentativo devido as suas semelhanças com a composição das outras comunidades analisadas. Espécies de Lactobacillus e Bacillus predominaram entre as amostras da primeira coleta. Halomonas, Streptococcus, Lactococcus e Pseudomonas foram detectados em amostras de biofilme e em amostras líquidas, sendo os principais contaminantes provindos de biofilme no momento da primeira coleta. Na segunda coleta Bacillus foi o principal contaminante e novamente gêneros produtores de ácido lático como Streptococcus, Lactobacillus e Staphylococcus foram os mais freqüentes. Os resultados concordam com o reportado na literatura sobre sistemas fermentativos convencionais. Apenas os primers desenhados para amplificação do gene 16S rRNA de Burkholderia, Pseudomonas e Weissella apresentaram especificidade em testes com isolados. Halomonas sp. foi encontrada em biofilme de dorna através do sequenciamento utilizando primers para esse gênero. Halomonas pode produzir levânio podendo haver o consumo da sacarose disponível para fermentação. Biofilme da centrífuga teve a maior quantidade de micro-organismos nos dois momentos de coleta (1,93E+06 UFC.mg-1 e 2,14E+07 UFC.mg-1, respectivamente) assim como as amostra de levedo entre as amostras líquidas (1,03E+08 UFC.ml-1 e 2,96E+06 UFC.ml-1, na primeira e segunda coleta, respectivamente), indicando níveis consideráveis de contaminantes. Burkholderia e Pseudomonas foram os mais abundantes entre as amostras de biofilmes da primeira e segunda coleta. Nas amostras líquidas Burkholderia apresentou-se em maior quantidade na maioria das amostras da primeira coleta; enquanto Pseudomonas e Weissella em geral predominaram equivalentemente entre as amostras da segunda coleta / Bacterial contamination by Lactobacillus, Bacillus and Leuconostoc and other lactic acid bacteria is one of the main factors that affects the yield in alcoholic fermentation process. Biofilm formation protects the bacteria community and it is a permanent source of contamination. For characterization of these contaminations in (1) biofilms from centrifuge, tank fermentation, heat exchanger and water pipe and (2) molasses, must, yeast, yeast treated (with H2SO4) and wine, samples were taken at two different periods from fermentation system characterized by high alcohol yields (16%). Restriction enzymes AluI, BstUI, HaeIII, HinfI, MseI and MspI used in T-RFLP analysis were defined by 16S rRNA gene sequences analysis in silico from common contaminants. These enzymes generate high number of unique T-RFs between 30 and 650 bp. DNA from samples were used as template in T-RFLP reactions in order to obtain molecular profiles of microbial communities present at each sample. Shannon diversity index was calculated based on T-RFs numbers. Principal component analysis (PCA) and phylogenetic inference of contaminants were performed based on T-RFs profiles. The main contaminant bacterial taxa were quantified by qPCR using specific primers designed in this study and considering the average of 16S rRNA gene copies previously counted into the genome of each bacterial taxon. Water pipe biofilm showed the highest rate of bacterial diversity in the samples collected in the first sampling period. For the samples collected in the second sampling, the highest rate of bacterial diversity was revealed for molasses and must. PCA suggested that biofilms (but not external sources) are the main contaminants in the studied fermentation process. It is probably due their similarities with the composition of other analyzed communities. Lactobacillus and Bacillus species predominated in first sampling period. Halomonas, Streptococcus, Lactococcus and Pseudomonas were detected in biofilm and liquid samples. They were the main contaminants from biofilm at this time of sampling. In the second sampling period, Bacillus was the most common genera and other lactic acid bacteria such Streptococcus, Staphylococcus and Lactobacillus were also the most frequent contaminants. These results agree with other reported in the literature about conventional fermentation systems. Only the primers designed in this study to amplify the 16S rRNA gene of Burkholderia, Pseudomonas and Weissella showed specificity in tests with bacterial strains. Halomonas sp. was revealed in biofilms from tank fermentation by DNA sequencing using designed primers for genera. Halomonas can produce levan and may consume sucrose available for generation of alcohol. Centrifugal biofilm had the highest amount of bacteria in both sampling periods (1.93E+06 CFU.mg-1 and 2.14E+07 CFU.mg-1, respectively). In liquid samples, yeast had the highest amount of bacteria in both sampling periods (1.03E+08 CFU.ml-1 and 2.96E+06 CFU.ml-1, respectively); it shows significant levels of contaminants. Burkholderia and Pseudomonas were more abundant among biofilm samples of all samplings. Burkholderia was present in high quantities in the majority of liquid samples taken during the first sampling period; Pseudomonas and Weissella equivalently predominated among samples taken during the second sampling period
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Avaliação da alterabilidade e da exalação do gás radônio em rochas ornamentais / Evaluation of alterability and exhalation os radon gas in dimension stonesAmaral, Paulo Giovany Quadros do 26 October 2016 (has links)
A presente tese se baseia no estudo das rochas ornamentais e de revestimento em relação aos seus aspectos de alterabilidade que irá influenciar diretamente no aumento da adição do gás radioativo radônio no interior de um ambiente com o passar do tempo de uso, fato devido as interações com o meio de onde é instalado a rocha, como exemplo da sua manutenção e limpeza, que implicará no seu padrão estético e na qualidade do ar do ambiente interior. Temas como a qualidade do ar interior relacionado a qualidade de vida do usuário, a radioatividade natural, o perigo do gás radiotivo radônio no interior do ambiente e as rochas ornamentais como composição e uso na arquitetura são apresentados para embasamento da Tese e auxiliando no entendimento dos ensaios realizados. Para isso, cinco rochas ornamentais foram escolhidas e realizados ensaios de alterabilidade química, ensaios tecnológicos de caraterização e ensaios de caracterização radiométrica a fim de prever o comportamento de uma dada rocha em seu ambiente ao ser aplicado na arquitetura. O ataque do ácido HCl nas rochas diminuiu sua resistência e aumentou a quantidade de radônio exalado relacionado diretamente ao aumento direto na porosidade, aumentando também a eficiência de exalação da rocha. O ataque do álcali KOH apesar de diminuir a resistência da rocha, interferiu na porosidade e diminuindo assim a quantidade de radônio exalado em alguns casos. Isso demonstra que mesmo conhecendo o material e a sua contribuição na adição do Rn no interior do ambiente, com o passar do tempo a rocha irá aumentar sua exalação quando não conservadas podendo aumentar o nível de radônio acumulado no ambiente. Com isso, a tese realça a importância de conhecer o material que irá compor o interior do ambiente afim de prevenir futuros danos à saúde da edificação e dos usuários, facilitando assim o entendimento desses materiais e como interfere na qualidade do ambiente construído. / This thesis is based on the study of dimension stone in relation to aspects of changeability which will directly influence the increased addition of radon radioactive gas within an environment over time of use, a fact due to interactions with the environment where it is installed rock, as an example of its maintenance and cleaning, which will result in its aesthetic standard and quality of indoor air environment. Topics such as the quality of the related indoor air to the user\'s quality of life, natural radioactivity, the danger of radiotivo radon gas into the atmosphere and dimension stone as composition and use in architecture are presented to foundation this Thesis and aiding in the understanding of tests. For this, five ornamental stones were chosen and performed chemical tests changeability, technological characterization of assays and radiometric characterization tests to predict the behavior of a given rock in your environment to be applied in architecture. The attack of the HCl acid in the rocks decreased its strength and increased the amount of exhaled radon related directly to the direct increase in porosity, also increasing exhalation efficiency of the rock. The attack KOH alkali spite of reducing the resistance of the rock, and interfere with the porosity thereby decreasing the amount of radon vented in some cases. This shows that even knowing the material and its contribution to the addition of Rn into the environment, with the passage of time the rock will increase your exhalation when not maintained can increase the level of accumulated radon in the environment. Thus, the thesis highlights the importance of knowing the material that will form the inside of the environment in order to prevent further damage to the health of the building and users, thus facilitating the understanding of these materials and how it interferes in the quality of the built environment.
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Determinação da distância de alcance dos pesticidas utilizados em forma de spray automático / Determinación de la distancia de alcance de los pesticidas utilizados em forma de spray automaticoRios, Eduardo Massoco 19 August 2017 (has links)
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Previous issue date: 2017-08-19 / Pelo uso crescente de inseticidas em residências há uma crescente preocupação com os efeitos na saúde da população pela utilização doméstica desses produtos. O consumidor, que na maioria das vezes, desconhece as propriedades tóxicas dos componentes dessas formulações como se fossem inócuos, porém na realidade foram encontrados sintomas como redução da performance intelectual, distúrbios visuais e auditivos, polineuropatia sensitivo-motora e alterações neurovegetativas, e foram encontradas evidências laboratoriais de distúrbios da imunidade celular e humoral pelo uso excessivo desses produtos. A piretrina, é um componente ativo extraído das flores de crisântemos e tem sido muito utilizada há mais de dois séculos no controle de inúmeras espécies de insetos. No Brasil tem seu uso autorizado para aplicação foliar nas culturas de algodão, arroz, café, couve, couve-flor, milho, repolho, soja, tomate e trigo. Os efeitos da exposição crônica aos contaminantes ambientais surgem após um intervalo de tempo variável podendo resultar em eventos adversos sobre o sistema nervoso, respiratório, cardiovascular e reprodutivo. Este trabalho teve como objetivo avaliar a dispersão do spray de pesticida residencial e sua maior concentração de piretróides em domicílios urbanos. Para a avaliação da propagação dos pesticidas em ambientes fechados foi realizado um teste com fórmula comercial para identificar o alcance da distância e a quantificação da concentração dos mesmos em cada um dos pontos. Foram 24 jatos em 4 horas. Nos ensaios de recuperação obteve-se valores entre 80 e 104% para as concentrações 1,3 e 6 mg.L-1. Encontrou-se uma concentração de 2,32 mg.L-1 para a menor distância da lata de spray e para a maior distância (11,4 m) a quantidade de pesticida encontrada foi de 0,58 mg.L-1. Isso demonstra que ao usar o dispositivo próximo aos móveis, poderá acarretar um risco maior de contaminação humana por pesticidas. Assim, estima-se que o uso indiscriminado de inseticidas nos domicílios cause danos na saúde humana, constituindo-se em um potencial problema de Saúde Pública. / There is a growing concern about the effects on the population health by the domestic use of these products due to the increasing use of insecticides in homes. The consumer, who for the most part is unaware of the toxic properties of the components of these formulations as if they were innocuous, however, symptoms such as reduced intellectual performance, visual and auditory disorders, sensory-motor polyneuropathy and neurovegetative alterations were found, also a laboratory evidence of cellular and humoral immunity observed disorders due to excessive use of these products. Pyrethrin is an active component extracted from chrysanthemum flowers and has been widely used for over two centuries in controlling numerous insect species. In Brazil, it has authorized its use for foliar application in cotton, rice, coffee, kale, cauliflower, corn, cabbage, soybean, tomato and wheat crops. The chronic exposure effects to environmental contaminants arise after a variable time interval and may result in adverse events on the nervous, respiratory, cardiovascular and reproductive systems. This work aimed to evaluate the dispersion of the domestic pesticide spray and its higher concentration of pyrethroids in urban households. For evaluation the pesticides spread indoors, a commercial pesticide test was performed to identify the range of pesticides and the extent of pesticide concentration at each point. There were twenty-four (24) jets in four hours. In the recovery essays were obtained values between 80 and 104% for 1,3 and 6 mg.L-1 concentrations. It was found a 2,32 mg.L-1 for the shortest distance from the spray and for the longest distance (11,4m) the amount of pesticide was 0,58 mg.L-1. It points out that using devices next to the furniture, it may carry a greater risk of human contamination by pesticides. Therefore, it is estimated that the indiscriminate use of insecticides in households causes damage to human health, constituting a potential public health problem. / Por el uso creciente de insecticidas en residencias hay uma creciente preocupación com los efectos en la salud de la población por la utilización doméstica de estos produtos. El consumidor que en la mayoría de las veces desconoce las propiedades toxicas de los componentes de esas fórmulas como si fuesen inofensivos. Pero en realidad se encontraron síntomas como reducción del comportamiento intelectual, disturbios visuales y auditivos, polineuropatías sensitiva motora y alteración neurovegetativas y fueron encontradas evidencias científicas de disturbios de la inmunidad celular y multicelular por el uso excessivo de estos productos. La piretrina, en un componente activo extraído de las flores del crisantemo y viene siendo utilizado hace más de dos siglos en el control de innumerables especies de insectos. En Brasil tiene su uso autorizado para la aplicación sobre las plantaciones de algodón, arroz, café, col, coliflor, maíz, repollo, soja, tomate, y trigo. Los efectos de la exposición crónica de los contaminantes ambientales surgen después de un intervalo de tiempo variable, pudiendo resultar en eventos adversos sobre el sistema nervioso, respiratorio, cardiovascular y/o reproductivo. Este trabajo tuvo como objetivo evaluar las aspersiones del spray del pesticida residencial y su mayor concentración de piretroides en domicilios urbanos. Para la evaluación de la propagación de los pesticidas en ambientes cerrados, fue realizado pruebas con una fórmula comercial para identificar el alcance de la distancia y la cantidad de cocentración de los mismos en cada uno de los puntos. Fueron 24 aspersiones en 4 horas. En los ensayos se obtuvo valores entre 80 y 104 % para las concentraciones 1, 3 y 6 mg.L-1. Se ha encontrado una concentración de 2,32 mg.L-1 para la menor distancia de la lata de spray y para mayor distancia (11,4 m) la cantidad de pesticida encontrado fue de 0,58 mg.L-1. Esto demuesta que al usar el dispositivo cerca de los muebles, puede acarrear un riesgo mayor de contaminación humana por pesticida. Así se estima que el uso indiscriminado de insecticidas en los domicilios cause daños en la salud humana, convirtiéndose en un potencial problema de salud pública.
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Characteristics and mechanisms of atrazine sorption to biochar for land remediationMcMillan, Oliver January 2018 (has links)
Contaminated land is a widespread, global issue affecting millions of people. Atrazine is a commonly used herbicide which often contaminates groundwater and drinking water supplies and is associated with adverse health outcomes. Biochar is the solid product of pyrolysis and is associated with several environmental benefits. It may be an effective remediation tool when used as a soil amendment. This thesis investigates the mechanisms through which biochar can immobilise atrazine, and the implications of the mechanisms for remediating contaminated land. Nine biochar samples were obtained from the United Kingdom Biochar Research Centre , which were produced from softwood pellets (SWP), wheat straw pellets (WSP), miscanthus straw pellets (MSP), rice husk (RH) and oil seed rape (OSR) each at pyrolysis temperatures of 550°C and 700°C (excluding OSR at 700°C). The sorption mechanisms controlling atrazine sorption to these biochars were determined through various characterisation methods and batch sorption experiments. The sorption tests showed that sorption to each of the standard biochars occurs via multiple simultaneously occurring mechanisms, which are each promoted under certain conditions. Studies investigating sorption kinetics, isotherms and interactions with humic acids showed that for all biochars in this study, pore filling was a significant process through which atrazine is transported to adsorption sites, although poor intraparticle diffusion for softwood and oil seed rape biochars can prevent efficient transport. Wheat straw and rice husk biochars showed effective pore diffusion, resulting in high sorption capacities. Partitioning was associated with poor remediation outcomes and was significant to softwood biochars, although adsorption dominated overall sorption for all other biochars. pH was shown to significantly influence the occurrence of various sorption mechanisms. At low pH values, most biochars showed evidence of electrostatic repulsion between positive atrazine species and the positively charged biochar surface. At intermediate pH values, all biochars showed strong hydrogen bonding between H+ groups on the surface of the biochar and atrazine. A meta-analysis of previous relevant studies provided further evidence for hydrogen bonding of atrazine to biochar and showed that hydrophobic effects likely play little role in adsorption after accounting for the effects of surface area. Varying contributions of π-π EDA interactions, hydrogen bonding involving biochar O- groups, and interactions with ash minerals resulted in different sorption profiles for each biochar at high pH values. In order to further determine the mechanisms controlling sorption at high pH, surface compositions of SWP550, RH700 and OSR550 biochars were modified using hydrofluoric acid. Modification with hydrofluoric acid successfully removed the ash contents of rice husk and oil seed rape biochars and reduced atrazine removal at high pH values. This suggested that the ash fraction increases atrazine removal at high pH through complexation or catalytic hydrolysis. The roles of the various mechanisms are related to remediation outcomes in a novel manner allowing for the improved design of biochar for environmental remediation.
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