South Africa ratified the Stockholm Convention (SC), which became legally binding on 17 May 2004. This Convention targets 12 particularly toxic persistent organic pollutants (POPs) for virtual elimination. The Convention also requires parties to reduce the release of organochlorine pesticides and the intentionally- and unintentionally-produced POPs such as dioxins, furans and polychlorinated biphenyls (PCBs) (referred to as dioxin-like chemicals). Dioxins are a heterogeneous mixture of chlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) congeners. These substances were never intentionally produced but are produced as by-products of industrial processes (such as metallurgical processes and bleaching of paper pulp). They can also be formed during natural processes such as volcanic eruptions and forest fires. The largest contributor to releases of PCDD/Fs in the environment is incomplete combustion from waste incinerators leading to the unintentional production of these compounds. Polychlorinated biphenyls (PCBs) are used in transformers and capacitors, but can also be formed unintentionally during industrial and thermal processes. Dioxin-like chemicals (PCDD/Fs and/or PCBs) are classified as persistent because of the following characteristics: lipophilicity and hydrophobicity; resistance to photolytic, chemical and biological degradation and they are able to travel long distances. As South Africa is a semiarid region, POPs will be less prone to travel here because these substances favour colder regions with high soil organic matter. Fish, predatory birds, mammals (including humans) absorb high concentrations of POPs through the process of bio-concentration, leading to bio-accumulation of these substances in the fatty tissue. PCDD/Fs occur as unwanted trace contaminants in air, water, land, in residues and products (such as consumer goods e.g. paper and textiles). The distribution of these chemicals into various matrices is problematic since they cause damage to the environment and human health. These chemicals pose a threat to human health when found in high concentrations that may lead to acute hepatoxicity and dermal toxicity (chloracne). Long-term exposure to low concentrations of these substances might lead to chronic effects such as reproductive problems and carcinogenicity. Since ferrous and non-ferrous metal production is a source of dioxin-like chemicals, a platinum mine in the Limpopo Province, South Africa, was selected for this investigation. The aim of the study was to determine if there are dioxin-like chemicals associated with platinum mining and processing, and if the H4IIE reporter gene bio-assay could be used to semi-quantify and assess the potencies of the complex environmental and process samples by determining their Toxic Equivalency Quotients (TEQ). The implications of the sources to the formation of dioxin-like chemicals regarding the SC were investigated and recommendations were made to improve this study. Samples were collected from tailings dams, woodchips, a dumpsite and slag from the smelter at Union Section. Samples were extracted with the Soxhlet apparatus using hexane as solvent. The percentage total organic carbon (%TOC) was determined for each sample to normalise the data. The method used was the Walkley-Black method. In determining the TEQ of each sample, the H4IIE luc cell line was used. The cells of the H4IIE luc line are genetically modified rat hepatoma cells stably transfected with a luciferase firefly gene. The luciferase gene is activated by the presence of dioxin-like compounds and the concentration of the enzyme is measured as relative light units (RLUs). The amount of RLUs is directly proportional to the dioxin load in the extract. This method is rapid, cost and time-effective in determining the TEQ when compared to chemical analysis. The TEQ2o-valuesin the various samples, as determined with the H4IIE luc cell line, ranged from 0.007 ngTEQ/kg to 54.06 ngTEQ/kg. Thermal processes at the smelter, sorption of hydrophobic organic compounds (HOCs) to soil and tailings, and external sources such as anthropogenic activities contributed to high TEQ2o-values. Climatic conditions, wind, precipitation, and solubility of HOCs into surfactants lead to low TEQ20. The smelter at Union Section had a very high TEQ20of 44.62 ngTEQ/kg compared to Impala Platinum mine (5.15 ngTEQ/kg). This implies that workers at Union Section are possibly exposed to low and high concentrations of dioxin-like chemicals. Long-term exposure to these compounds could lead to bio-accumulation in the fatty tissue of the mine workers, leading to chronic effects such as reproductive problems and cancer. The air emission of the furnace at the smelter was 0.03 gTEQ/annum and the release of the PCDD/Fs into the slag was 0.60 gTEQ/annum. By effectively managing the smelter it is possible to reduce the TEQ. The TEQ of each sample increased due to normalising the data. The normalised TEQ20 ranged from 0.94 ng TEQ/kg to 42497.48 ngTEQ/kg. Dioxin-like chemicals are present on a platinum mine, but at varying quantities and the effects of these compounds might be detrimental to the environment and the workers at the platinum mine. Further analyses of the health impacts associated with the platinum mine are needed. The H4IIE reporter gene bio-assay could be used to effectively determine the TEQ of each sample. Although this investigation has identified the formation and presence of dioxin-like chemicals at certain stages of mining and processing, not all of the processes were investigated. Some of these processes have the potential to add, and even destroy, these chemicals, affecting potential human exposure and amounts released to the environment. This, however, requires further investigation. The financial assistance of the National Research Foundation (NRF) towards this research is hereby acknowledged. Opinions expressed and conclusions arrived at, are those of the author and are not necessarily to be attributed to the NRF. / Thesis (M. Environmental Science)--North-West University, Potchefstroom Campus, 2006.

Farmed Southern Bluefin Tuna (SBT) (Thunnus maccoyii) is an important export product for South Australia (SA). It is exported to Japan, China, Korea and the United States for the sushi and sashimi markets. The primary purpose of SBT farming in SA is to fatten wild-caught juvenile fish (2-4 years of age with initial mean weights between 12-20 kg) over a period of approximately five months by feeding a selection of baitfish types. Farmers, farm managers and consumers of SBT all have an interest in managing chemical residues that have the potential to biomagnify in the fatty tissue of the farmed SBT fillets. Of particular interest are chemical residues of polychlorinated biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins / dibenzofurans (PCDD/Fs). This research presents the investigations and experimental validation of a predictive model that can be used to address the levels of residues in the fillets of farmed SBT at harvest from feeding (as the source) when applied to SBT aquaculture. An additional industry-focussed aim of this research was to determine if a Longer Term Holding (LTH) farming period, with a duration of an extra 12 months after a typical farming period of approximately five months, could produce SBT with higher condition index (CI) and lipid content, while keeping levels of PCBs and PCDD/Fs low, compared to the typical farming period. The justification for this research is that an adequate quantitative model is essential to help industry achieve targeted concentrations in the final fillet product by making scientific-based decisions on baitfish selection (baitfish strategies for the feeding of SBT), and longer term, to confidently demonstrate to local markets and importing countries that Australia is actively managing levels of PCBs and PCDD/Fs in farmed SBT, to ensure a high quality and safe product is delivered to the consumer. The novelty of this research is underpinned by four integrated stages, and the criteria for an adequate model established. The important criteria included: accurate predictions versus observed data demonstrated through the analysis of residual plots, potential physiological interpretation of model coefficients, parsimony – the model should be as simple as possible (but no simpler) and that the model should be easy to use. Firstly, a logical starting point was the development of a risk framework for residues in SBT. The developed framework was based on conventional principles of microbiological risk assessment highlighted in Codex Alimentarius. The risk framework consists of five governing principles: hazard identification, hazard characterisation, exposure assessment, risk characterisation and model validation. The advantages of the risk framework is that it provides a systematic research approach and permits information to be handled unambiguously, especially important for the niche SBT industry where chemical residue research is carried out for the first time. Secondly, because of a lack of available scientific data in context of this research, commercial-scale experimental field data for levels of PCBs and PCDD/Fs in typical farmed SBT as affected by feeding and growth were collected over 17 months at seven time intervals from Farm Delta Fishing Pty Ltd in 2005/06 (n = 50). Field data from another commercial company, Farm Alpha Fishing Pty Ltd, was collected over the typical farming period specific to this company, spanning 15 weeks at three time intervals in 2006, for validation work (n = 15). The data obtained from Farm Delta Fishing Pty Ltd revealed that whole weight of farmed SBT increased from 18.5 kg to 30.3 kg for a typical farming period, and subsequently to 41.0 kg by the end of the LTH farming period. A maximum mean CI of 24.0 ± 0.5 kg.m⁻³ and a maximum mean lipid content of 17.6 ± 0.5% was achieved at the third time interval of the typical farming period, for the baitfish types and ratios used as feed. There were no significant differences in the CI and lipid between the final harvests of the typical farming and LTH periods, i.e. even after an additional 12 months of farming. PCB and PCDD/F concentrations, however, increased between the final harvests of the typical farming and LTH periods. The data indicated that a typical farming period was sufficient to achieve a maximum CI and lipid content with lower concentrations of PCBs and PCDD/Fs in the fillets relative to the LTH farming period. For the third stage of this research, a quantitative model was synthesised and applied to the PCB and PCDD/F (2,3,7,8-TeCDF) data detected in farmed SBT fillets. Assimilation efficiencies for PCBs and 2,3,7,8-TeCDF in the fillets of SBT were obtained. An assimilation efficiency, or percentage retention (efficiency expressed as a percentage), in the fillet of SBT is a measure of the uptake of a chemical residue from food (baitfish) to the SBT fillet. For the WHO-PCBs, assimilation efficiencies based on SBT fillets ranged between 19.1 – 35.3 % with the exception of PCB 169. The highest assimilation efficiency of 35.3 %, with a range of 30.4 – 40.3 % (at the 95 % confidence level) was attributed to the most toxic PCB congener, PCB 126. An assimilation efficiency of 39.2 % was determined in SBT fillets for the congener 2,3,7,8-TeCDF, which was higher than the assimilation efficiencies determined for the WHO-PCB congeners. A residual plot as predicted value versus observed value indicated that the predictive model was neither under- or over-parameterised. However, when the predictive model was assessed against the data set from Farm Alpha Pty Ltd, the model over-predicted the actual PCB and PCDD/F concentrations. The over-prediction is attributed to possible overfeeding of SBT farmed by Farm Alpha Fishing Pty Ltd. From a food safety point of view, in the absence of ideal predictions because of a lack of ideal validation data sets, an over-prediction instead of under-prediction is preferred. In the fourth stage, the practical application of the predictive model was demonstrated. Because SBT fillets are retailed as tissue group-specific, i.e. akami (low fat), chu-toro (medium fat) and otoro (high fat) fillets, PCB and PCDD/F analyses were carried out on the three tissue groups for selected SBT (n = 7). Dietary modelling on SBT consumption in humans was carried out using findings from the predictive model and tissue-specific data. The baitfish strategy employed for the feeding of farmed SBT consequently affects dietary exposure to SBT consumers. Exposure to PCBs and PCDD/Fs is approximately seven times lower for the consumption of a skin-free, boneless akami fillet than for a comparable otoro fillet of the same size. This dietary exposure assessment accounted only for consumption of SBT tissue-specific fillets. The experimental field study and modelling work on PCB and PCDD/F concentrations in farmed SBT (fillets) outlined in this thesis importantly directs the need to re-evaluate a specific model to better cater for SBT farming practices where SBT fillets are produced for human consumption. Because conditions that normally pertain to commercial farming of wild-caught fish were studied, findings should be of interest to industries where other species of fish (for food) are farmed in sea-cages in the open ocean. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1342453 / Thesis (Ph.D.) -- University of Adelaide, School of Chemical Engineering, 2008

Farmed Southern Bluefin Tuna (SBT) (Thunnus maccoyii) is an important export product for South Australia (SA). It is exported to Japan, China, Korea and the United States for the sushi and sashimi markets. The primary purpose of SBT farming in SA is to fatten wild-caught juvenile fish (2-4 years of age with initial mean weights between 12-20 kg) over a period of approximately five months by feeding a selection of baitfish types. Farmers, farm managers and consumers of SBT all have an interest in managing chemical residues that have the potential to biomagnify in the fatty tissue of the farmed SBT fillets. Of particular interest are chemical residues of polychlorinated biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins / dibenzofurans (PCDD/Fs). This research presents the investigations and experimental validation of a predictive model that can be used to address the levels of residues in the fillets of farmed SBT at harvest from feeding (as the source) when applied to SBT aquaculture. An additional industry-focussed aim of this research was to determine if a Longer Term Holding (LTH) farming period, with a duration of an extra 12 months after a typical farming period of approximately five months, could produce SBT with higher condition index (CI) and lipid content, while keeping levels of PCBs and PCDD/Fs low, compared to the typical farming period. The justification for this research is that an adequate quantitative model is essential to help industry achieve targeted concentrations in the final fillet product by making scientific-based decisions on baitfish selection (baitfish strategies for the feeding of SBT), and longer term, to confidently demonstrate to local markets and importing countries that Australia is actively managing levels of PCBs and PCDD/Fs in farmed SBT, to ensure a high quality and safe product is delivered to the consumer. The novelty of this research is underpinned by four integrated stages, and the criteria for an adequate model established. The important criteria included: accurate predictions versus observed data demonstrated through the analysis of residual plots, potential physiological interpretation of model coefficients, parsimony – the model should be as simple as possible (but no simpler) and that the model should be easy to use. Firstly, a logical starting point was the development of a risk framework for residues in SBT. The developed framework was based on conventional principles of microbiological risk assessment highlighted in Codex Alimentarius. The risk framework consists of five governing principles: hazard identification, hazard characterisation, exposure assessment, risk characterisation and model validation. The advantages of the risk framework is that it provides a systematic research approach and permits information to be handled unambiguously, especially important for the niche SBT industry where chemical residue research is carried out for the first time. Secondly, because of a lack of available scientific data in context of this research, commercial-scale experimental field data for levels of PCBs and PCDD/Fs in typical farmed SBT as affected by feeding and growth were collected over 17 months at seven time intervals from Farm Delta Fishing Pty Ltd in 2005/06 (n = 50). Field data from another commercial company, Farm Alpha Fishing Pty Ltd, was collected over the typical farming period specific to this company, spanning 15 weeks at three time intervals in 2006, for validation work (n = 15). The data obtained from Farm Delta Fishing Pty Ltd revealed that whole weight of farmed SBT increased from 18.5 kg to 30.3 kg for a typical farming period, and subsequently to 41.0 kg by the end of the LTH farming period. A maximum mean CI of 24.0 ± 0.5 kg.m⁻³ and a maximum mean lipid content of 17.6 ± 0.5% was achieved at the third time interval of the typical farming period, for the baitfish types and ratios used as feed. There were no significant differences in the CI and lipid between the final harvests of the typical farming and LTH periods, i.e. even after an additional 12 months of farming. PCB and PCDD/F concentrations, however, increased between the final harvests of the typical farming and LTH periods. The data indicated that a typical farming period was sufficient to achieve a maximum CI and lipid content with lower concentrations of PCBs and PCDD/Fs in the fillets relative to the LTH farming period. For the third stage of this research, a quantitative model was synthesised and applied to the PCB and PCDD/F (2,3,7,8-TeCDF) data detected in farmed SBT fillets. Assimilation efficiencies for PCBs and 2,3,7,8-TeCDF in the fillets of SBT were obtained. An assimilation efficiency, or percentage retention (efficiency expressed as a percentage), in the fillet of SBT is a measure of the uptake of a chemical residue from food (baitfish) to the SBT fillet. For the WHO-PCBs, assimilation efficiencies based on SBT fillets ranged between 19.1 – 35.3 % with the exception of PCB 169. The highest assimilation efficiency of 35.3 %, with a range of 30.4 – 40.3 % (at the 95 % confidence level) was attributed to the most toxic PCB congener, PCB 126. An assimilation efficiency of 39.2 % was determined in SBT fillets for the congener 2,3,7,8-TeCDF, which was higher than the assimilation efficiencies determined for the WHO-PCB congeners. A residual plot as predicted value versus observed value indicated that the predictive model was neither under- or over-parameterised. However, when the predictive model was assessed against the data set from Farm Alpha Pty Ltd, the model over-predicted the actual PCB and PCDD/F concentrations. The over-prediction is attributed to possible overfeeding of SBT farmed by Farm Alpha Fishing Pty Ltd. From a food safety point of view, in the absence of ideal predictions because of a lack of ideal validation data sets, an over-prediction instead of under-prediction is preferred. In the fourth stage, the practical application of the predictive model was demonstrated. Because SBT fillets are retailed as tissue group-specific, i.e. akami (low fat), chu-toro (medium fat) and otoro (high fat) fillets, PCB and PCDD/F analyses were carried out on the three tissue groups for selected SBT (n = 7). Dietary modelling on SBT consumption in humans was carried out using findings from the predictive model and tissue-specific data. The baitfish strategy employed for the feeding of farmed SBT consequently affects dietary exposure to SBT consumers. Exposure to PCBs and PCDD/Fs is approximately seven times lower for the consumption of a skin-free, boneless akami fillet than for a comparable otoro fillet of the same size. This dietary exposure assessment accounted only for consumption of SBT tissue-specific fillets. The experimental field study and modelling work on PCB and PCDD/F concentrations in farmed SBT (fillets) outlined in this thesis importantly directs the need to re-evaluate a specific model to better cater for SBT farming practices where SBT fillets are produced for human consumption. Because conditions that normally pertain to commercial farming of wild-caught fish were studied, findings should be of interest to industries where other species of fish (for food) are farmed in sea-cages in the open ocean. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1342453 / Thesis (Ph.D.) -- University of Adelaide, School of Chemical Engineering, 2008

Farmed Southern Bluefin Tuna (SBT) (Thunnus maccoyii) is an important export product for South Australia (SA). It is exported to Japan, China, Korea and the United States for the sushi and sashimi markets. The primary purpose of SBT farming in SA is to fatten wild-caught juvenile fish (2-4 years of age with initial mean weights between 12-20 kg) over a period of approximately five months by feeding a selection of baitfish types. Farmers, farm managers and consumers of SBT all have an interest in managing chemical residues that have the potential to biomagnify in the fatty tissue of the farmed SBT fillets. Of particular interest are chemical residues of polychlorinated biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins / dibenzofurans (PCDD/Fs). This research presents the investigations and experimental validation of a predictive model that can be used to address the levels of residues in the fillets of farmed SBT at harvest from feeding (as the source) when applied to SBT aquaculture. An additional industry-focussed aim of this research was to determine if a Longer Term Holding (LTH) farming period, with a duration of an extra 12 months after a typical farming period of approximately five months, could produce SBT with higher condition index (CI) and lipid content, while keeping levels of PCBs and PCDD/Fs low, compared to the typical farming period. The justification for this research is that an adequate quantitative model is essential to help industry achieve targeted concentrations in the final fillet product by making scientific-based decisions on baitfish selection (baitfish strategies for the feeding of SBT), and longer term, to confidently demonstrate to local markets and importing countries that Australia is actively managing levels of PCBs and PCDD/Fs in farmed SBT, to ensure a high quality and safe product is delivered to the consumer. The novelty of this research is underpinned by four integrated stages, and the criteria for an adequate model established. The important criteria included: accurate predictions versus observed data demonstrated through the analysis of residual plots, potential physiological interpretation of model coefficients, parsimony – the model should be as simple as possible (but no simpler) and that the model should be easy to use. Firstly, a logical starting point was the development of a risk framework for residues in SBT. The developed framework was based on conventional principles of microbiological risk assessment highlighted in Codex Alimentarius. The risk framework consists of five governing principles: hazard identification, hazard characterisation, exposure assessment, risk characterisation and model validation. The advantages of the risk framework is that it provides a systematic research approach and permits information to be handled unambiguously, especially important for the niche SBT industry where chemical residue research is carried out for the first time. Secondly, because of a lack of available scientific data in context of this research, commercial-scale experimental field data for levels of PCBs and PCDD/Fs in typical farmed SBT as affected by feeding and growth were collected over 17 months at seven time intervals from Farm Delta Fishing Pty Ltd in 2005/06 (n = 50). Field data from another commercial company, Farm Alpha Fishing Pty Ltd, was collected over the typical farming period specific to this company, spanning 15 weeks at three time intervals in 2006, for validation work (n = 15). The data obtained from Farm Delta Fishing Pty Ltd revealed that whole weight of farmed SBT increased from 18.5 kg to 30.3 kg for a typical farming period, and subsequently to 41.0 kg by the end of the LTH farming period. A maximum mean CI of 24.0 ± 0.5 kg.m⁻³ and a maximum mean lipid content of 17.6 ± 0.5% was achieved at the third time interval of the typical farming period, for the baitfish types and ratios used as feed. There were no significant differences in the CI and lipid between the final harvests of the typical farming and LTH periods, i.e. even after an additional 12 months of farming. PCB and PCDD/F concentrations, however, increased between the final harvests of the typical farming and LTH periods. The data indicated that a typical farming period was sufficient to achieve a maximum CI and lipid content with lower concentrations of PCBs and PCDD/Fs in the fillets relative to the LTH farming period. For the third stage of this research, a quantitative model was synthesised and applied to the PCB and PCDD/F (2,3,7,8-TeCDF) data detected in farmed SBT fillets. Assimilation efficiencies for PCBs and 2,3,7,8-TeCDF in the fillets of SBT were obtained. An assimilation efficiency, or percentage retention (efficiency expressed as a percentage), in the fillet of SBT is a measure of the uptake of a chemical residue from food (baitfish) to the SBT fillet. For the WHO-PCBs, assimilation efficiencies based on SBT fillets ranged between 19.1 – 35.3 % with the exception of PCB 169. The highest assimilation efficiency of 35.3 %, with a range of 30.4 – 40.3 % (at the 95 % confidence level) was attributed to the most toxic PCB congener, PCB 126. An assimilation efficiency of 39.2 % was determined in SBT fillets for the congener 2,3,7,8-TeCDF, which was higher than the assimilation efficiencies determined for the WHO-PCB congeners. A residual plot as predicted value versus observed value indicated that the predictive model was neither under- or over-parameterised. However, when the predictive model was assessed against the data set from Farm Alpha Pty Ltd, the model over-predicted the actual PCB and PCDD/F concentrations. The over-prediction is attributed to possible overfeeding of SBT farmed by Farm Alpha Fishing Pty Ltd. From a food safety point of view, in the absence of ideal predictions because of a lack of ideal validation data sets, an over-prediction instead of under-prediction is preferred. In the fourth stage, the practical application of the predictive model was demonstrated. Because SBT fillets are retailed as tissue group-specific, i.e. akami (low fat), chu-toro (medium fat) and otoro (high fat) fillets, PCB and PCDD/F analyses were carried out on the three tissue groups for selected SBT (n = 7). Dietary modelling on SBT consumption in humans was carried out using findings from the predictive model and tissue-specific data. The baitfish strategy employed for the feeding of farmed SBT consequently affects dietary exposure to SBT consumers. Exposure to PCBs and PCDD/Fs is approximately seven times lower for the consumption of a skin-free, boneless akami fillet than for a comparable otoro fillet of the same size. This dietary exposure assessment accounted only for consumption of SBT tissue-specific fillets. The experimental field study and modelling work on PCB and PCDD/F concentrations in farmed SBT (fillets) outlined in this thesis importantly directs the need to re-evaluate a specific model to better cater for SBT farming practices where SBT fillets are produced for human consumption. Because conditions that normally pertain to commercial farming of wild-caught fish were studied, findings should be of interest to industries where other species of fish (for food) are farmed in sea-cages in the open ocean. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1342453 / Thesis (Ph.D.) -- University of Adelaide, School of Chemical Engineering, 2008

A emissão de uma grande diversidade de substâncias através dos efluentes industriais gasosos, líquidos ou sólidos acarreta que uma gama de compostos seja introduzida no ambiente atmosférico e aquático de meios urbanos. A exposição a substâncias genotóxicas no ambiente oferece freqüentemente um grande risco à saúde humana. Com essa preocupação, o presente trabalho avaliou a exposição das pessoas à qualidade do ar atmosférico e à da água para abastecimento público de duas cidades urbanas do Rio Grande do Sul (Brasil), que recebem diferentes contribuições antrópicas (cidades-alvo), comparando-as com uma cidade menos impactada - cidade referência. Os extratos orgânicos da água tratada para abastecimento público e das amostras de ar foram testados quanto a mutagenicidade através do ensaio Salmonella/microssoma (linhagens TA98 e TA100 para água e ar, com e sem fração de metabolização e, ainda para ar, YG1021 e YG1024) - marcador de mutagenicidade. As amostras de ar ainda foram analisadas por HPLC para a caracterização de HPAs e algumas amostras também por HRGC-HRMS para a composição dos PCDD/Fs. Para o biomonitoramento humano foram avaliados através do ensaio do cometa e do micronúcleo (MN), em linfócitos e células da mucosa bucal, respectivamente, as pessoas que viviam nas três cidades - biomarcador de efeito. Análises de polimorfismos genéticos (CYP1A1, GSTM1 e GSTT1) foram utilizados como biomarcadores de susceptibilidade. As duas cidades-alvo apresentaram resultados significativos quanto a mutagenicidade da água de abastecimento apresentando todos os resultados positivos, principalmente para mutação de substituição de pares de bases (TA100), indicando a presença de compostos como THMs e MX. Valores de PTS atmosféricos apresentaram-se em algumas amostragens acima dos limites da legislação brasileira (CONAMA, 1990). Os HPAs encontrados em maior quantidade foram indeno(1,2,3-c,d)pireno e benzo(ghi)perileno, sendo que em uma das amostras a análise dos PCDD/Fs, mostrou valores que superaram em dez vezes a outra amostra analisada. As duas cidades alvo apresentaram altas respostas mutagênicas nas amostras de ar tanto por mutágenos de ação direta, como indireta e também altas concentrações de nitro-HPAs. Estes resultados sinalizaram para a necessidade de uma revisão da legislação para material particulado atmosférico, já que dentro dos atuais parâmetros, grande quantidade de compostos mutagênicos foram encontrados. Para os indivíduos estudados quanto aos biomarcadores de efeito, tanto os linfócitos analisados pelo ensaio do cometa quanto às células da mucosa bucal pelo ensaio do MN não apresentaram resultados significativos, sendo que apenas uma cidade alvo apresentou quanto ao ensaio do cometa. Como a maioria dos indivíduos CYP1A1, GSTM1 e GSTT1 apresentaram genótipos prevalentes, pode estar ocorrendo uma eficiência no sistema de detoxificação garantidos pela presença desses genótipos. Este trabalho mostrou a importância de estudos ambientais que relacionem marcadores de mutagenicidade com biomarcadores de efeito e de susceptibilidade, para que haja uma maior compreensão dos efeitos da exposição humana diária à agentes genotóxicos ambientais. / The emission of a great substances amount through gaseous, liquid and solid industrial effluents have been introduced many kinds of chemical compounds in aquatic and atmospheric environments. The environmental exposure to genotoxic substances often brings great risk to human health. Therefore, the aim of this work was make an avaliation of people exposition to atmospheric air and water supply quality in two urban cities in Rio Grande do Sul State (Brazil) which were under influence of different anthropogenic sources (target cities), compared to a less intense influenced urban city (reference city). The organic extracts from supply water and airborne samples were tested to mutagenicity using the Salmonella/microsome assay (TA98 and TA100 strains for water and airborne samples, with and without metabolization fraction, and also, YG1021 and YG1024 strains for airborne) - mutagenicity marker. Moreover, the airborne samples were analysed by HPLC to PAHs composition and by HRGC-HRMS to PCDD/Fs composition. Human biomonitoring was evaluated by samples of lymphocytes and bucal mucosa cells from individuals living in this three cities using the comet and the micronuclei (MN) assays, respectively - effect biomarkers. Analysis of gene polymorphisms (CYP1A1, GSTM1 and GSTT1) was taken as susceptibility biomarkers. Both target cities showed significative results for the water mutagenicity with mutagenic responses for all samples, mainly to base-pair substitution mutation (TA100 strain), denoting the presence of THMs and MX compounds. The TSP concentrations were above the Brazilian Legislation and World Health Organization regulation limits few times. The PAHs detected in greater amout were indeno(1,2,3-c,d)pyrene and benzo(ghi)perylene and one of the samples overcame ten times the values of PCDD/Fs compared to the other analysed. The two target cities showed high mutagenicity response in all airborne samples, both by direct and indirect mutagens, besides high concentrations of nitro-PHAs. Actually, these results indicate a law revision necessity, because great amount of mutagenic compounds were assessed in smaller concentration that those present in current legal parameters. The analysed subjects for effect biomarkers, both limphocytes tested by comet assay and buccal mucosa cells tested by MN, did not show significant results, only one city show increased primary DNA damage detected through comet assay. As the majority of subjects CYP1A1, GSTM1 e GSTT1 showed prevalent genotype, it maybe occuring an efficient system detoxification warranted by these genotypes presence. This work has shown the importance of environmental studies that relate mutagenicity markers with effect and susceptibility biomarkers, thus a greater understanding about environmental genotoxic agents, which people are exposed daily, will be done carefully.

A emissão de uma grande diversidade de substâncias através dos efluentes industriais gasosos, líquidos ou sólidos acarreta que uma gama de compostos seja introduzida no ambiente atmosférico e aquático de meios urbanos. A exposição a substâncias genotóxicas no ambiente oferece freqüentemente um grande risco à saúde humana. Com essa preocupação, o presente trabalho avaliou a exposição das pessoas à qualidade do ar atmosférico e à da água para abastecimento público de duas cidades urbanas do Rio Grande do Sul (Brasil), que recebem diferentes contribuições antrópicas (cidades-alvo), comparando-as com uma cidade menos impactada - cidade referência. Os extratos orgânicos da água tratada para abastecimento público e das amostras de ar foram testados quanto a mutagenicidade através do ensaio Salmonella/microssoma (linhagens TA98 e TA100 para água e ar, com e sem fração de metabolização e, ainda para ar, YG1021 e YG1024) - marcador de mutagenicidade. As amostras de ar ainda foram analisadas por HPLC para a caracterização de HPAs e algumas amostras também por HRGC-HRMS para a composição dos PCDD/Fs. Para o biomonitoramento humano foram avaliados através do ensaio do cometa e do micronúcleo (MN), em linfócitos e células da mucosa bucal, respectivamente, as pessoas que viviam nas três cidades - biomarcador de efeito. Análises de polimorfismos genéticos (CYP1A1, GSTM1 e GSTT1) foram utilizados como biomarcadores de susceptibilidade. As duas cidades-alvo apresentaram resultados significativos quanto a mutagenicidade da água de abastecimento apresentando todos os resultados positivos, principalmente para mutação de substituição de pares de bases (TA100), indicando a presença de compostos como THMs e MX. Valores de PTS atmosféricos apresentaram-se em algumas amostragens acima dos limites da legislação brasileira (CONAMA, 1990). Os HPAs encontrados em maior quantidade foram indeno(1,2,3-c,d)pireno e benzo(ghi)perileno, sendo que em uma das amostras a análise dos PCDD/Fs, mostrou valores que superaram em dez vezes a outra amostra analisada. As duas cidades alvo apresentaram altas respostas mutagênicas nas amostras de ar tanto por mutágenos de ação direta, como indireta e também altas concentrações de nitro-HPAs. Estes resultados sinalizaram para a necessidade de uma revisão da legislação para material particulado atmosférico, já que dentro dos atuais parâmetros, grande quantidade de compostos mutagênicos foram encontrados. Para os indivíduos estudados quanto aos biomarcadores de efeito, tanto os linfócitos analisados pelo ensaio do cometa quanto às células da mucosa bucal pelo ensaio do MN não apresentaram resultados significativos, sendo que apenas uma cidade alvo apresentou quanto ao ensaio do cometa. Como a maioria dos indivíduos CYP1A1, GSTM1 e GSTT1 apresentaram genótipos prevalentes, pode estar ocorrendo uma eficiência no sistema de detoxificação garantidos pela presença desses genótipos. Este trabalho mostrou a importância de estudos ambientais que relacionem marcadores de mutagenicidade com biomarcadores de efeito e de susceptibilidade, para que haja uma maior compreensão dos efeitos da exposição humana diária à agentes genotóxicos ambientais. / The emission of a great substances amount through gaseous, liquid and solid industrial effluents have been introduced many kinds of chemical compounds in aquatic and atmospheric environments. The environmental exposure to genotoxic substances often brings great risk to human health. Therefore, the aim of this work was make an avaliation of people exposition to atmospheric air and water supply quality in two urban cities in Rio Grande do Sul State (Brazil) which were under influence of different anthropogenic sources (target cities), compared to a less intense influenced urban city (reference city). The organic extracts from supply water and airborne samples were tested to mutagenicity using the Salmonella/microsome assay (TA98 and TA100 strains for water and airborne samples, with and without metabolization fraction, and also, YG1021 and YG1024 strains for airborne) - mutagenicity marker. Moreover, the airborne samples were analysed by HPLC to PAHs composition and by HRGC-HRMS to PCDD/Fs composition. Human biomonitoring was evaluated by samples of lymphocytes and bucal mucosa cells from individuals living in this three cities using the comet and the micronuclei (MN) assays, respectively - effect biomarkers. Analysis of gene polymorphisms (CYP1A1, GSTM1 and GSTT1) was taken as susceptibility biomarkers. Both target cities showed significative results for the water mutagenicity with mutagenic responses for all samples, mainly to base-pair substitution mutation (TA100 strain), denoting the presence of THMs and MX compounds. The TSP concentrations were above the Brazilian Legislation and World Health Organization regulation limits few times. The PAHs detected in greater amout were indeno(1,2,3-c,d)pyrene and benzo(ghi)perylene and one of the samples overcame ten times the values of PCDD/Fs compared to the other analysed. The two target cities showed high mutagenicity response in all airborne samples, both by direct and indirect mutagens, besides high concentrations of nitro-PHAs. Actually, these results indicate a law revision necessity, because great amount of mutagenic compounds were assessed in smaller concentration that those present in current legal parameters. The analysed subjects for effect biomarkers, both limphocytes tested by comet assay and buccal mucosa cells tested by MN, did not show significant results, only one city show increased primary DNA damage detected through comet assay. As the majority of subjects CYP1A1, GSTM1 e GSTT1 showed prevalent genotype, it maybe occuring an efficient system detoxification warranted by these genotypes presence. This work has shown the importance of environmental studies that relate mutagenicity markers with effect and susceptibility biomarkers, thus a greater understanding about environmental genotoxic agents, which people are exposed daily, will be done carefully.

A emissão de uma grande diversidade de substâncias através dos efluentes industriais gasosos, líquidos ou sólidos acarreta que uma gama de compostos seja introduzida no ambiente atmosférico e aquático de meios urbanos. A exposição a substâncias genotóxicas no ambiente oferece freqüentemente um grande risco à saúde humana. Com essa preocupação, o presente trabalho avaliou a exposição das pessoas à qualidade do ar atmosférico e à da água para abastecimento público de duas cidades urbanas do Rio Grande do Sul (Brasil), que recebem diferentes contribuições antrópicas (cidades-alvo), comparando-as com uma cidade menos impactada - cidade referência. Os extratos orgânicos da água tratada para abastecimento público e das amostras de ar foram testados quanto a mutagenicidade através do ensaio Salmonella/microssoma (linhagens TA98 e TA100 para água e ar, com e sem fração de metabolização e, ainda para ar, YG1021 e YG1024) - marcador de mutagenicidade. As amostras de ar ainda foram analisadas por HPLC para a caracterização de HPAs e algumas amostras também por HRGC-HRMS para a composição dos PCDD/Fs. Para o biomonitoramento humano foram avaliados através do ensaio do cometa e do micronúcleo (MN), em linfócitos e células da mucosa bucal, respectivamente, as pessoas que viviam nas três cidades - biomarcador de efeito. Análises de polimorfismos genéticos (CYP1A1, GSTM1 e GSTT1) foram utilizados como biomarcadores de susceptibilidade. As duas cidades-alvo apresentaram resultados significativos quanto a mutagenicidade da água de abastecimento apresentando todos os resultados positivos, principalmente para mutação de substituição de pares de bases (TA100), indicando a presença de compostos como THMs e MX. Valores de PTS atmosféricos apresentaram-se em algumas amostragens acima dos limites da legislação brasileira (CONAMA, 1990). Os HPAs encontrados em maior quantidade foram indeno(1,2,3-c,d)pireno e benzo(ghi)perileno, sendo que em uma das amostras a análise dos PCDD/Fs, mostrou valores que superaram em dez vezes a outra amostra analisada. As duas cidades alvo apresentaram altas respostas mutagênicas nas amostras de ar tanto por mutágenos de ação direta, como indireta e também altas concentrações de nitro-HPAs. Estes resultados sinalizaram para a necessidade de uma revisão da legislação para material particulado atmosférico, já que dentro dos atuais parâmetros, grande quantidade de compostos mutagênicos foram encontrados. Para os indivíduos estudados quanto aos biomarcadores de efeito, tanto os linfócitos analisados pelo ensaio do cometa quanto às células da mucosa bucal pelo ensaio do MN não apresentaram resultados significativos, sendo que apenas uma cidade alvo apresentou quanto ao ensaio do cometa. Como a maioria dos indivíduos CYP1A1, GSTM1 e GSTT1 apresentaram genótipos prevalentes, pode estar ocorrendo uma eficiência no sistema de detoxificação garantidos pela presença desses genótipos. Este trabalho mostrou a importância de estudos ambientais que relacionem marcadores de mutagenicidade com biomarcadores de efeito e de susceptibilidade, para que haja uma maior compreensão dos efeitos da exposição humana diária à agentes genotóxicos ambientais. / The emission of a great substances amount through gaseous, liquid and solid industrial effluents have been introduced many kinds of chemical compounds in aquatic and atmospheric environments. The environmental exposure to genotoxic substances often brings great risk to human health. Therefore, the aim of this work was make an avaliation of people exposition to atmospheric air and water supply quality in two urban cities in Rio Grande do Sul State (Brazil) which were under influence of different anthropogenic sources (target cities), compared to a less intense influenced urban city (reference city). The organic extracts from supply water and airborne samples were tested to mutagenicity using the Salmonella/microsome assay (TA98 and TA100 strains for water and airborne samples, with and without metabolization fraction, and also, YG1021 and YG1024 strains for airborne) - mutagenicity marker. Moreover, the airborne samples were analysed by HPLC to PAHs composition and by HRGC-HRMS to PCDD/Fs composition. Human biomonitoring was evaluated by samples of lymphocytes and bucal mucosa cells from individuals living in this three cities using the comet and the micronuclei (MN) assays, respectively - effect biomarkers. Analysis of gene polymorphisms (CYP1A1, GSTM1 and GSTT1) was taken as susceptibility biomarkers. Both target cities showed significative results for the water mutagenicity with mutagenic responses for all samples, mainly to base-pair substitution mutation (TA100 strain), denoting the presence of THMs and MX compounds. The TSP concentrations were above the Brazilian Legislation and World Health Organization regulation limits few times. The PAHs detected in greater amout were indeno(1,2,3-c,d)pyrene and benzo(ghi)perylene and one of the samples overcame ten times the values of PCDD/Fs compared to the other analysed. The two target cities showed high mutagenicity response in all airborne samples, both by direct and indirect mutagens, besides high concentrations of nitro-PHAs. Actually, these results indicate a law revision necessity, because great amount of mutagenic compounds were assessed in smaller concentration that those present in current legal parameters. The analysed subjects for effect biomarkers, both limphocytes tested by comet assay and buccal mucosa cells tested by MN, did not show significant results, only one city show increased primary DNA damage detected through comet assay. As the majority of subjects CYP1A1, GSTM1 e GSTT1 showed prevalent genotype, it maybe occuring an efficient system detoxification warranted by these genotypes presence. This work has shown the importance of environmental studies that relate mutagenicity markers with effect and susceptibility biomarkers, thus a greater understanding about environmental genotoxic agents, which people are exposed daily, will be done carefully.

The recent developments and improvements of instrumental methods for the analyses of the environmental contaminants, especially the persistent organic pollutants (POPs), have made it possible to detect and quantify these at very low concentrations in environmental and biotic matrices. The main objective of this thesis is to demonstrate the capability of the atmospheric pressure chemical ionization technique (APCI), using gas chromatography coupled to tandem mass spectrometry for the determination of a wide range of environmental contaminants, including the POPs regulated by Stockholm Convention, such as polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), but also the derivates of PBDEs and novel brominated flame retardants (NBFRs). The APCI was operated in charge transfer condition, preferably producing molecular ions. Multiple reaction monitoring (MRM) experiments were optimized by adjusting cone voltage, collision energy and dwell time. Optimization of source parameters, such as gas flows and temperatures was also performed. Low concentration standards were analyzed, achieving a visible chromatographic peak for 2 fg 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) demonstrating the excellent sensitivity of the system. Adequate linearity and repeatability were observed for all the studied compounds. The performance of APCI methods was validated against the conventional methods using gas chromatography coupled to high resolution mass spectrometry for chlorinated compounds in a wide range of matrices including environmental, air, human and food matrices. The GC-APCI-MS/MS method was successfully applied to a set of 75 human serum samples to study the circulating levels of POPs in epidemiologic studies. Moreover the method was utilized to establish temporal trends of POPs in osprey eggs samples collected during the past five decades.

Polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs), two groups of structurally related chlorinated aromatic hydrocarbons, are of high concern due to their global distribution and extreme toxicity. Since they occur at very low levels, their analysis is complex, challenging and hence there is a need for efficient, reliable and rapid alternative analytical methods. Developing such methods was the aim of the project this thesis is based upon. During the first years of the project the focus was on the first parts of the analytical chain (extraction and clean-up). A selective pressurized liquid extraction (SPLE) procedure was developed, involving in-cell clean-up to remove bulk co-extracted matrix components from sample extracts. It was further streamlined by employing a modular pressurized liquid extraction (M-PLE) system, which simultaneously extracts, cleans up and isolates planar PCDD/Fs in a single step. Both methods were validated using a wide range of soil, sediment and sludge reference materials. Using dichloromethane/n-heptane (DCM/Hp; 1/1, v/v) as a solvent, results statistically equivalent to or higher than the reference values were obtained, while an alternative, less harmful non-chlorinated solvent mixture - diethyl ether/n-heptane (DEE/Hp; 1/2, v/v) – yielded data equivalent to those values. Later, the focus of the work shifted to the final instrumental analysis. Six gas chromatography (GC) phases were evaluated with respect to their chromatographic separation of not just the 17 most toxic congeners (2,3,7,8-substituted PCDD/Fs), but all 136 tetra- to octaCDD/Fs. Three novel ionic liquid columns performed much better than previously tested commercially available columns. Supelco SLB-IL61 offered the best overall performance, successfully resolving 106 out of the 136 compounds, and 16 out of the 17 2,3,7,8-substituted PCDD/Fs. Another ionic liquid (SLB-IL111) column provided complementary separation. Together, the two columns separated 128 congeners. The work also included characterization of 22 GC columns’ selectivity and solute-stationary phase interactions. The selectivities were mapped using Principal Component Analysis (PCA) of all 136 PCDD/F’s retention times on the columns, while the interactions were probed by analyzing both the retention times and the substances’ physicochemical properties.

Persistent organic pollutants (POPs) are hydrophobic substances that readily sorb to organic matter in particles and colloids instead of being freely dissolved in the water phase. This sorption affects the bio­availability and environmental transport of the POPs. The major part of this thesis concerns the role of sediments as secondary sources of POPs. As the primary emissions decrease, contaminated sediments where POPs have accumulated can become the main source of contamination. If the contaminated sediment by time becomes covered with cleaner layers, the POPs are buried and no longer in contact with the aquatic environment. Experiments in this thesis showed, however, that new invading species can alter the sediment-water dynamics as a result of their bioturbation, i.e. mixing of sediment particles and pore-water. Marenzelleria spp., invading species in the Baltic Sea that burrow deeper than native species, were found to increase the remobilization of buried contaminants. The sediment-to-water flux was inversely related to the burial depth (2-10 cm) of the POP congeners (polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers) and also inversely related to the hydrophobicity of the congener. The flux was therefore most pronounced for less hydrophobic contaminants, which was linked to the bioirrigating behaviour of these species. Marenzelleria spp. also accumulated the buried POPs and increased concentrations in surface sedi­ment. Contaminants previously considered buried at a ’safe’ depth can thus be remobilized as a result of the invasion of Marenzelleria spp. in the Baltic Sea. One method to decrease the remobilization of contaminants from sediments is ’capping’, i.e. a layer of clean material is placed as a cap on the sediment. By amending the cap with active materials, which sequester the POPs and decrease their availability, thinner layers can be used (’active capping’ or ’thin-layer capping’). Results from an experiment with thin-layer capping using different active materials (activated carbon (AC) and kraft lignin) showed that both the sediment-to-water flux and the bioaccumulation by benthic species of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), hexachlorobenzene (HCB) and octachlorostyrene (OCS) decreased with increased thick­ness of the cap layer (0.5-5 cm). Amendments with active materials further increased the cap efficiency. AC was more efficient than kraft lignin, and a 3 cm cap with 3.3% AC reduced the flux and bioaccumulation with ~90%. The reduction of the sediment-to-water flux was inversely related to the hydrophobicity of the POP, and reductions in the flux had similar magnitudes as reductions in the concentration in deep-burrowing polychaetes, demonstrating the importance of bioturbation for sediment-to-water transport. In a one-year study on the levels of PCDD/Fs, PCBs, and HCB in a coastal area of the Baltic Sea, the correlations between the POP levels and the levels of particles and organic carbon in the water were found to differ for POPs of different structure and hydrophobicity. The levels of PCDD/Fs decreased to one third in May, which could be related to the increased sedimentation, i.e. water-to-sediment transport, during spring bloom.