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The screening of phyto-pesticides for potential adverse effects on human healthShoko, Yeukai Phoebe January 2010 (has links)
<p>Pesticides are designed to control or eliminate pests such as insects, rodents, weeds,<br />
bacteria, and fungi. They are used at a global scale for agricultural produce. Although<br />
pesticides play a significant role in increasing food production and eliminating diseases,<br />
exposure to pesticides may be harmful to non-target organisms. As a result concern over<br />
safety and resistance to pesticides has increased and there is pressure to reduce use and<br />
search for more environmentally and toxicologically safe and efficacious pesticides. Most<br />
pesticides currently in use are synthetic / therefore an alternative to synthetic pesticides is<br />
the use of naturally occurring products/ botanicals with pesticidal properties.</p>
<p>Two plants indigenous to South African with pesticidal properties were chosen for this<br />
study. Dicerothamnus rhinocerotis (D. rhinocerotis) and Galenia africana (G. africana)<br />
have potential antifungal properties thus, may have potential use on agricultural produce<br />
as fungicides. Galenia africana and D. rhinocerotis extracts inhibit growth of B. cinerea<br />
(a fungal pathogen) at concentrations greater than 31.25 mg/ml and 125mg/ml<br />
respectively. A major consideration in approving pesticides for use is whether they pose<br />
an unreasonable risk to humans and to the environment. Toxicity studies are required to<br />
determine the safety of the plant extracts.</p>
<p>The purpose of this study was to evaluate potential toxicity of ethanol extracts of D.<br />
rhinocerotis and G. africana, which is important when designing practices to reduce or<br />
eliminate excess exposure to them. Natural plant products with pesticidal properties could<br />
provide an alternative to synthetic pesticides and may thus effectively reduce resistance<br />
levels.<br />
<br />
<br />
  / </p>
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The screening of phyto-pesticides for potential adverse effects on human healthShoko, Yeukai Phoebe January 2010 (has links)
<p>Pesticides are designed to control or eliminate pests such as insects, rodents, weeds,<br />
bacteria, and fungi. They are used at a global scale for agricultural produce. Although<br />
pesticides play a significant role in increasing food production and eliminating diseases,<br />
exposure to pesticides may be harmful to non-target organisms. As a result concern over<br />
safety and resistance to pesticides has increased and there is pressure to reduce use and<br />
search for more environmentally and toxicologically safe and efficacious pesticides. Most<br />
pesticides currently in use are synthetic / therefore an alternative to synthetic pesticides is<br />
the use of naturally occurring products/ botanicals with pesticidal properties.</p>
<p>Two plants indigenous to South African with pesticidal properties were chosen for this<br />
study. Dicerothamnus rhinocerotis (D. rhinocerotis) and Galenia africana (G. africana)<br />
have potential antifungal properties thus, may have potential use on agricultural produce<br />
as fungicides. Galenia africana and D. rhinocerotis extracts inhibit growth of B. cinerea<br />
(a fungal pathogen) at concentrations greater than 31.25 mg/ml and 125mg/ml<br />
respectively. A major consideration in approving pesticides for use is whether they pose<br />
an unreasonable risk to humans and to the environment. Toxicity studies are required to<br />
determine the safety of the plant extracts.</p>
<p>The purpose of this study was to evaluate potential toxicity of ethanol extracts of D.<br />
rhinocerotis and G. africana, which is important when designing practices to reduce or<br />
eliminate excess exposure to them. Natural plant products with pesticidal properties could<br />
provide an alternative to synthetic pesticides and may thus effectively reduce resistance<br />
levels.<br />
<br />
<br />
  / </p>
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The screening of phyto-pesticides for potential adverse effects on human healthShoko, Yeukai Phoebe January 2010 (has links)
Philosophiae Doctor - PhD / Pesticides are designed to control or eliminate pests such as insects, rodents, weeds, bacteria, and fungi. They are used at a global scale for agricultural produce. Although pesticides play a significant role in increasing food production and eliminating diseases, exposure to pesticides may be harmful to non-target organisms. As a result concern over safety and esistance to pesticides has increased and there is pressure to reduce use and search for more environmentally and toxicologically safe and efficacious pesticides. Most pesticides currently in use are synthetic; therefore an alternative to synthetic pesticides is the use of naturally occurring products/ botanicals with pesticidal properties. Two plants indigenous to South African with pesticidal properties were chosen for this study. Dicerothamnus rhinocerotis (D. rhinocerotis) and Galenia africana (G. africana) have potential antifungal properties thus, may have potential use on agricultural produce as fungicides. Galenia africana and D. rhinocerotis extracts inhibit growth of B. cinerea (a fungal pathogen) at concentrations greater than 31.25 mg/ml and 125mg/ml respectively. A major consideration in approving pesticides for use is whether they pose
an unreasonable risk to humans and to the environment. Toxicity studies are required to determine the safety of the plant extracts.
The purpose of this study was to evaluate potential toxicity of ethanol extracts of D. rhinocerotis and G. africana, which is important when designing practices to reduce or eliminate excess exposure to them. Natural plant products with pesticidal properties could provide an alternative to synthetic pesticides and may thus effectively reduce resistance levels. This first objective of this study was to assess the cytotoxicity of D. rhinocerotis and G. africana on human cell cultures. Human whole blood and the human breast adenocarcinoma cell line (MCF-7) were treated with varying concentrations of the plant extracts and cytotoxicity determined. Cytotoxicity was measured using several biomarkers. Inhibiotory concentration for a 50% effect (IC50) and no observable effect
level (NOEL) values were obtained for D. rhinocerotis and G. africana. The results showed that extracts of D. rhinocerotis and G. africana had cytotoxic effects on the cell cultures. The second objective of this study was to determine the ecotoxicity of D. rhinocerotis and G. africana. A series of acute toxicity tests, with effective concentration for a 50% effect (EC50) and lethal concentration for a 50% effect (LC50) as biomarkers, were
conducted to estimate the potential environmental effect of the two plants. The tests were carried out using Vibrio fischeri, Selenastrum capricornutum, Daphnia pulex, and Poecilia reticulata as bioindicators. Results obtained showed that G. africana had higher toxicity units than D. rhinocerotis, thus showing that G. africana is more toxic to the aquatic species used as compared to D. rhinocerotis. The third objective of this study was to investigate the immunomodulatory effects of the two plant extracts. This was achieved by using mouse spleen cell cultures. Exposure of pesticides to the immune system may result in alteration of the normal immune functions. The cytokines IFN-γ and IL-4 were used as biomarkers to determine the T-cell activity of the immune system when exposed to the two botanical products. The results obtained showed that both D. rhinocerotis and G. africana decrease of the cytokines interferongamma
(IFN-γ) and interleukin-4 (IL-4), thus may have immunotoxic effects.
The fourth objective was to investigate the hepatotoxicity of the two plant extracts. Injury to the liver was investigated using a range of clinical biochemical tests that monitor liver enzyme activity and metabolic activity. Primary liver cell cultures were exposed to the plant products in question, after which the biochemical tests were carried out. The enzymes that were monitored were alanine aminotrasferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH). The results obtained showed that both of D. rhinocerotis and G. africana may have effects on the liver, as shown by the increased levels of enzymes released from cells upon exposure to plant extracts. The final objective of this study was to investigate the effect of the two plants on the male reproductive system. Injury to the male reproductive system was investigated using testicular cell cultures. Primary cell cultures were stimulated with luteinizing-hormone (LH) and exposed to the plant extracts. LH results in the production of testosterone, thus
testosterone was used as a biomarker for assessing reproductive toxicity. The results obtained showed that both of D. rhinocerotis and G. africana have effects on the male reproductive system, as shown by the decreased testosterone secretion. Botanicals provide a simple, inexpensive and environmentally friendly (non-pollution and lesser toxicological concerns) alternative for pest control. However, motivation for the commercial use of botanicals as pesticides requires validating the efficacy of the plant as a pesticide, and also assessing its effects on human health and the environment. An important component of this evaluation involves toxicity studies, which enables cautions of dangerous practices and toxic effects of the plants to be issued. / South Africa
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The screening of phyto-pesticides for potential adverse effects on human healthShoko, Yeukai Phoebe January 2010 (has links)
Philosophiae Doctor - PhD / Pesticides are designed to control or eliminate pests such as insects, rodents, weeds, bacteria, and fungi. They are used at a global scale for agricultural produce. Although pesticides playa significant role in increasing food production and eliminating diseases, exposure to pesticides may be harmful to non-target organisms. As a result concern over safety and resistance to pesticides has increased and there is pressure to reduce use and search for more environmentally and toxicologically safe and efficacious pesticides. Most pesticides currently in use are synthetic; therefore an alternative to synthetic pesticides is the use of naturally occurring products! botanicals with pesticidal properties.
Two plants indigenous to South African with pesticidal properties were chosen for this study. Dicerothamnus rhinocerotis (D. rhinocerotis) and Galenia africana (G. africana) have potential antifungal properties thus, may have potential use on agricultural produce as fungicides. Galenia africana and D. rhinocerotis extracts inhibit growth of B. cinerea (a fungal pathogen) at concentrations greater than 31.25 mg/ml and 125mg/ml respectively. A major consideration in approving pesticides for use is whether they pose an unreasonable risk to humans and to the environment. Toxicity studies are required to determine the safety of the plant extracts. The purpose of this study was to evaluate potential toxicity of ethanol extracts of D. rhinocerotis and G. africana, which is important when designing practices to reduce or eliminate excess exposure to them. Natural plant products with pesticidal properties could provide an alternative to synthetic pesticides and may thus effectively reduce resistance levels. This first objective of this study was to assess the cytotoxicity of D. rhinocerotis and G. africana on human cell cultures. Human whole blood and the human breast adenocarcinoma cell line (MCF-7) were treated with varying concentrations of the plant extracts and cytotoxicity determined. Cytotoxicity was measured using several biomarkers. Inhibiotory concentration for a 50% effect (ICso) and no observable effect level (NOEL) values were obtained for D. rhinocerotis and G. africana. The results showed that extracts of D. rhinocerotis and G. africana had cytotoxic effects on the cell cultures. The second objective of this study was to determine the ecotoxicity of D. rhinocerotis and G. africana. A series of acute toxicity tests, with effective concentration for a 50% effect (ECso) and lethal concentration for a 50% effect (LCso) as biomarkers, were conducted to estimate the potential environmental effect of the two plants. The tests were
carried out using Vibrio fischeri, Selenastrum capricornutum, Daphnia pulex, and Poecilia reticulata as bioindicators. Results obtained showed that G. africana had higher toxicity units than D. rhinocerotis, thus showing that G. africana is more toxic to the aquatic species used as compared to D. rhinocerotis. The third objective of this study was to investigate the immunomodulatory effects of the two plant extracts. This was achieved by using mouse spleen cell cultures. Exposure of pesticides to the immune system may result in alteration of the normal immune functions. The cytokines IFN-y and IL-4 were used as biomarkers to determine the T-cell activity of the immune system when exposed to the two botanical products. The results obtained showed that both D. rhinocerotis and G. africana decrease of the cytokines interferon gamma (IFN-y) and interleukin-4 (IL-4), thus may have immunotoxic effects. The fourth objective was to investigate the hepatotoxicity of the two plant extracts. Injury to the liver was investigated using a range of clinical biochemical tests that monitor liver enzyme activity and metabolic activity. Primary liver cell cultures were exposed to the plant products in question, after which the biochemical tests were carried
out. The enzymes that were monitored were alanine aminotrasferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH). The results obtained showed that both of D. rhinocerotis and G. africana may have effects on the liver, as shown by the increased levels of enzymes released from cells upon exposure to plant extracts. The final objective of this study was to investigate the effect of the two plants on the male reproductive system. Injury to the male reproductive system was investigated using
testicular cell cultures. Primary cell cultures were stimulated with luteinizing-hormone (LH) and exposed to the plant extracts. LH results in the production of testosterone, thus testosterone was used as a biomarker for assessing reproductive toxicity. The results obtained showed that both of D. rhinocerotis and G. africana have effects on the male reproductive system, as shown by the decreased testosterone secretion. Botanicals provide a simple, inexpensive and environmentally friendly (non-pollution and lesser toxicological concerns) alternative for pest control. However, motivation for the commercial use of botanicals as pesticides requires validating the efficacy of the plant as a pesticide, and also assessing its effects on human health and the environment. An important component of this evaluation involves toxicity studies, which enables cautions of dangerous practices and toxic effects of the plants to be issued.
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Efeitos da exposição in vivo à hidroquinona sobre funções do tecido traqueal e de macrófagos alveolares de camundongos / Effects of in vivo hydroquinone exposure on functions of alveolar macrophages and tracheal tissue in miceShimada, Ana Lucia Borges 08 April 2011 (has links)
A hidroquinona (HQ) é um composto fenólico de origem natural ou antropogênica, encontrada em grandes concentrações no cigarro, além de ser produto da biotransformação do benzeno. Nosso grupo de pesquisa tem demonstrado que a exposição à HQ compromete a resposta inflamatória in vivo. Dando continuidade a estas investigações, este trabalho visou investigar os efeitos da exposição in vivo à HQ sobre funções do tecido traqueal e atividades de macrófagos alveolares. Para tanto, camundongos Swiss machos foram expostos à HQ 25ppm (1,5mg/60mL/1h; 5 dias) ou veículo (solução salina etanol 1:20) por via sistêmica (nebulização). Concentrações de mediadores inflamatórios (interleucina (IL) 1β, IL-6, IL-10, IL-4, IL-12 fator de necrose tumoral-α (TNF-α) ou proteína quimiotáxica de monócitos (MCP-1); ensaio imunoenzimático (ELISA) e óxido nítrico (NO; reação de Griess) foram quantificados no lavado bronco-alveolar (LBA) em condições basais ou 3 horas após estímulo inflamatório (LPS in vivo; 100µL/mL; 10min) e no sobrenadante de macrófagos (MΦs) alveolares ou de cultura da traquéia obtidos dos animais e posteriormente estimulados in vitro (MΦs: 5µg/mL de LPS + 10ng/mL de IFN-γ; traquéia: 1µg/mL de LPS; 24h). Atividades fagocítica e fungicida (microscopia óptica) e a expressão de receptores envolvidos na fagocitose toll-like receptor (TLR, TLR-2, TLR4 e dectina-1; citometria de fluxo) foram determinadas após incubação in vitro de MΦs alveolares com o fungo Candida albicans e a expressão protéica de MyD88 foi realizada em MΦs alveolares (western blot). Quantificação do RNAm para MCP-1 (reação da transcriptase reversa em cadeia de polimerase, RT-PCR) foi realizada em tecido traqueal e células de linhagem monocítica humana THP-1 foram empregadas em ensaios de quimiotaxia in vitro (Câmara de Boyden) frente a diferentes concentrações de MCP-1. Reatividade do tecido traqueal foi quantificada frente à metacolina. Os resultados obtidos mostraram que a exposição in vivo à HQ reduziu a concentração de MCP-1 (54,98% vs. controle LPS) e IL-12 (51,45% vs. controle LPS) no LBA após inflamação; reduziu a secreção de MCP-1 por MΦs (basal: 87,96%; LPS+INF-γ: 61,20%) e tecido traqueal em cultura (79,77% vs. controle LPS). Neste último tecido, a diminuição foi dependente da menor expressão gênica. Concentrações de MCP-1 semelhantes à detectada no sobrenadante de cultura de traquéia de animais expostos à HQ induziram migração de células THP-1 menor (38,2%) que à provocada pela concentração de MCP-1 no sobrenadante da cultura traquéia de animais controles. Traquéias de animais expostos à HQ apresentaram hiperreatividade (193,48%), a qual foi revertida pela remoção do epitélio traqueal. Adicionalmente, cultura de MΦs obtidos de animais expostos à HQ apresentaram maior atividade fagocítica (porcentagem de fagocitose: 36,30%; índice de fagocitose: 83,97%) e fungicida (68,47%), que não foram dependentes de alterações nos receptores TLR2, TLR4 e dectina-1, mas podem ser decorrentes da menor expressão protéica de MyD88. Em conjunto, os dados obtidos apontam para alterações importantes resultantes da exposição in vivo à HQ sobre funções de MΦs alveolares e do tecido traqueal que, em conjunto, podem ser determinantes para a toxicidade observada nestes animais que culmina com prejuízo na defesa do organismo. / Hydroquinone (HQ) is a phenolic compound of natural or anthropogenic source, also found in high concentrations in cigarette, as well as benzene´s metabolite. Our research group has demonstrated that exposure to HQ impairs in vivo inflammatory response. Following these investigations, this work aimed to study the effects of in vivo exposure to HQ on tracheal tissue and alveolar macrophages (MΦs) activities. For this purpose, male Swiss mice were systemically (aerolised) exposed to 25ppm HQ (1.5 mg/60mL/1h; 5 days) or vehicle (saline ethanol solution, 1:20). Concentrations of inflammatory mediators (interleukin (IL) IL-1β, IL- 6, IL-10, IL-4, IL-12 tumor necrosis factor-α (TNF-α ) or monocyte chemoattractant protein (MCP-1); (enzyme immune assay, ELISA)) and nitric oxide (NO; Griess reaction) were quantified in bronchoalveolar lavage (BAL) at baseline or 3 hours after inflammatory stimulus (in vivo LPS, 100µL/mL; 10min); in the supernatant of cultured alveolar macrophages (MΦs) or trachea obtained from animals and subsequently in vitro stimulated (MΦs: 5µg/mL of LPS plus 10ng/mL IFN-γ; trachea: 1µg/mL LPS; 24 hours). Phagocytic and fungicidal activities (light microscopy) and expression of receptors involved in phagocytosis (toll-like receptor (TLR, TLR2, TLR4 and dectin-1, flow cytometry) were determined after in vitro incubation of alveolar MΦs with Candida albicans fungus and expression of MyD88 pathway was held in alveolar MΦs (western blot). Quantification of mRNA for MCP-1 (reaction of reverse transcriptase polymerase chain reaction, RT-PCR) was performed in tracheal tissue and cells human monocytic THP-1 were used in in vitro chemotaxis assays (Boyden chamber) using different concentrations of MCP-1. Tracheal reactivity was measured in response to methacholine. The results showed that in vivo HQ exposure reduced the concentration of MCP-1 (54.98% vs. control) and IL-12 (51.45% vs. control) in the BAL after inflammation; decreased secretion of MCP-1 by MΦs (basal: 87.96%, LPS+INF-γ: 61.20%) and in tracheal culture after LPS stimulation (79.77% vs. control). In the latter tissue, the MCP-1 protein content was dependent on impaired gene expression. Concentrations of MCP-1 similar to those detected in the supernatant of tracheal from HQ exposed rats induced smaller migration of THP-1 cells (38.2%) than that evoked by the MCP-1 concentration obtained in trachea supernatants collected from control animals. Tracheas from HQ exposed rats showed hyper reactivity (193.48%), which was reversed by removal of the tracheal epithelium. Additionally, culture of MΦs obtained from HQ exposed rats showed increased phagocytic (percentage of phagocytosis: 36.30%; phagocytosis index: 83.97%) and fungicide activity (68.47%), which were not dependent on changes in the receptors TLR2, TLR4 and dectin-1, but could be due to reduced MyD88 expression. Together, these data point out important alterations on MΦs and trachea after in vivo HQ exposure, which may be crucial for the toxicity observed in these animals that culminates with impaired host defense.
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Efeitos da exposição in vivo à hidroquinona sobre funções do tecido traqueal e de macrófagos alveolares de camundongos / Effects of in vivo hydroquinone exposure on functions of alveolar macrophages and tracheal tissue in miceAna Lucia Borges Shimada 08 April 2011 (has links)
A hidroquinona (HQ) é um composto fenólico de origem natural ou antropogênica, encontrada em grandes concentrações no cigarro, além de ser produto da biotransformação do benzeno. Nosso grupo de pesquisa tem demonstrado que a exposição à HQ compromete a resposta inflamatória in vivo. Dando continuidade a estas investigações, este trabalho visou investigar os efeitos da exposição in vivo à HQ sobre funções do tecido traqueal e atividades de macrófagos alveolares. Para tanto, camundongos Swiss machos foram expostos à HQ 25ppm (1,5mg/60mL/1h; 5 dias) ou veículo (solução salina etanol 1:20) por via sistêmica (nebulização). Concentrações de mediadores inflamatórios (interleucina (IL) 1β, IL-6, IL-10, IL-4, IL-12 fator de necrose tumoral-α (TNF-α) ou proteína quimiotáxica de monócitos (MCP-1); ensaio imunoenzimático (ELISA) e óxido nítrico (NO; reação de Griess) foram quantificados no lavado bronco-alveolar (LBA) em condições basais ou 3 horas após estímulo inflamatório (LPS in vivo; 100µL/mL; 10min) e no sobrenadante de macrófagos (MΦs) alveolares ou de cultura da traquéia obtidos dos animais e posteriormente estimulados in vitro (MΦs: 5µg/mL de LPS + 10ng/mL de IFN-γ; traquéia: 1µg/mL de LPS; 24h). Atividades fagocítica e fungicida (microscopia óptica) e a expressão de receptores envolvidos na fagocitose toll-like receptor (TLR, TLR-2, TLR4 e dectina-1; citometria de fluxo) foram determinadas após incubação in vitro de MΦs alveolares com o fungo Candida albicans e a expressão protéica de MyD88 foi realizada em MΦs alveolares (western blot). Quantificação do RNAm para MCP-1 (reação da transcriptase reversa em cadeia de polimerase, RT-PCR) foi realizada em tecido traqueal e células de linhagem monocítica humana THP-1 foram empregadas em ensaios de quimiotaxia in vitro (Câmara de Boyden) frente a diferentes concentrações de MCP-1. Reatividade do tecido traqueal foi quantificada frente à metacolina. Os resultados obtidos mostraram que a exposição in vivo à HQ reduziu a concentração de MCP-1 (54,98% vs. controle LPS) e IL-12 (51,45% vs. controle LPS) no LBA após inflamação; reduziu a secreção de MCP-1 por MΦs (basal: 87,96%; LPS+INF-γ: 61,20%) e tecido traqueal em cultura (79,77% vs. controle LPS). Neste último tecido, a diminuição foi dependente da menor expressão gênica. Concentrações de MCP-1 semelhantes à detectada no sobrenadante de cultura de traquéia de animais expostos à HQ induziram migração de células THP-1 menor (38,2%) que à provocada pela concentração de MCP-1 no sobrenadante da cultura traquéia de animais controles. Traquéias de animais expostos à HQ apresentaram hiperreatividade (193,48%), a qual foi revertida pela remoção do epitélio traqueal. Adicionalmente, cultura de MΦs obtidos de animais expostos à HQ apresentaram maior atividade fagocítica (porcentagem de fagocitose: 36,30%; índice de fagocitose: 83,97%) e fungicida (68,47%), que não foram dependentes de alterações nos receptores TLR2, TLR4 e dectina-1, mas podem ser decorrentes da menor expressão protéica de MyD88. Em conjunto, os dados obtidos apontam para alterações importantes resultantes da exposição in vivo à HQ sobre funções de MΦs alveolares e do tecido traqueal que, em conjunto, podem ser determinantes para a toxicidade observada nestes animais que culmina com prejuízo na defesa do organismo. / Hydroquinone (HQ) is a phenolic compound of natural or anthropogenic source, also found in high concentrations in cigarette, as well as benzene´s metabolite. Our research group has demonstrated that exposure to HQ impairs in vivo inflammatory response. Following these investigations, this work aimed to study the effects of in vivo exposure to HQ on tracheal tissue and alveolar macrophages (MΦs) activities. For this purpose, male Swiss mice were systemically (aerolised) exposed to 25ppm HQ (1.5 mg/60mL/1h; 5 days) or vehicle (saline ethanol solution, 1:20). Concentrations of inflammatory mediators (interleukin (IL) IL-1β, IL- 6, IL-10, IL-4, IL-12 tumor necrosis factor-α (TNF-α ) or monocyte chemoattractant protein (MCP-1); (enzyme immune assay, ELISA)) and nitric oxide (NO; Griess reaction) were quantified in bronchoalveolar lavage (BAL) at baseline or 3 hours after inflammatory stimulus (in vivo LPS, 100µL/mL; 10min); in the supernatant of cultured alveolar macrophages (MΦs) or trachea obtained from animals and subsequently in vitro stimulated (MΦs: 5µg/mL of LPS plus 10ng/mL IFN-γ; trachea: 1µg/mL LPS; 24 hours). Phagocytic and fungicidal activities (light microscopy) and expression of receptors involved in phagocytosis (toll-like receptor (TLR, TLR2, TLR4 and dectin-1, flow cytometry) were determined after in vitro incubation of alveolar MΦs with Candida albicans fungus and expression of MyD88 pathway was held in alveolar MΦs (western blot). Quantification of mRNA for MCP-1 (reaction of reverse transcriptase polymerase chain reaction, RT-PCR) was performed in tracheal tissue and cells human monocytic THP-1 were used in in vitro chemotaxis assays (Boyden chamber) using different concentrations of MCP-1. Tracheal reactivity was measured in response to methacholine. The results showed that in vivo HQ exposure reduced the concentration of MCP-1 (54.98% vs. control) and IL-12 (51.45% vs. control) in the BAL after inflammation; decreased secretion of MCP-1 by MΦs (basal: 87.96%, LPS+INF-γ: 61.20%) and in tracheal culture after LPS stimulation (79.77% vs. control). In the latter tissue, the MCP-1 protein content was dependent on impaired gene expression. Concentrations of MCP-1 similar to those detected in the supernatant of tracheal from HQ exposed rats induced smaller migration of THP-1 cells (38.2%) than that evoked by the MCP-1 concentration obtained in trachea supernatants collected from control animals. Tracheas from HQ exposed rats showed hyper reactivity (193.48%), which was reversed by removal of the tracheal epithelium. Additionally, culture of MΦs obtained from HQ exposed rats showed increased phagocytic (percentage of phagocytosis: 36.30%; phagocytosis index: 83.97%) and fungicide activity (68.47%), which were not dependent on changes in the receptors TLR2, TLR4 and dectin-1, but could be due to reduced MyD88 expression. Together, these data point out important alterations on MΦs and trachea after in vivo HQ exposure, which may be crucial for the toxicity observed in these animals that culminates with impaired host defense.
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Assessment of embryotoxicity of the antiandrogenic drugs flutamide and bicalutamide in zebrafish (Danio rerio)Holmlund, Josefin January 2020 (has links)
Introduction: Prostate cancer is the most common type of cancer in Sweden and is often treated using antiandrogenic drug therapy. Two substances belonging to this class of pharmaceuticals are bicalutamide and flutamide. After excretion from the human body, the drug molecules enter the wastewater treatment plant (WWTP). The WWTPs are not effective enough to completely remove pharmaceutical residues, why presence of both bicalutamide and flutamide can be detected in WWTP effluent water. Previous findings: Antiandrogens have been reported to affect reproduction in adult fish, but studies regarding possible effects on the embryonic development of fish are few. Aim: The present study sought to investigate if exposure to bicalutamide or flutamide cause toxicity in the early developmental stages of zebrafish embryos, and whether negative effects occur within concentrations relevant to measured environmental levels. Method: A modified OECD FET-test was used, where additional sublethal endpoints were included and the time period for assessment extended to 144 hours post fertilization (hpf). In addition, a locomotor activity assay was performed at 144 hpf in order to observe any sub-lethal swimming behavioral effects. Results: High doses (10 mg/L) of flutamide led to 100% lethality of the zebrafish embryos but the results suggest no acute toxic effects in the high dose treatment group of bicalutamide, or of either flutamide or bicalutamide within in the low (0.1 mg/L) or intermediate (1 mg/L) treatment groups. Neither did the locomotor activity assay result in statistically significant results, although the pattern of swimming activity in the low dose groups suggests that behavioral developmental effects could be present. Conclusions: High doses of flutamide caused mortality of the embryos, but no lethal or sublethal effects were present at environmentally relevant concentrations. The modest outcome of present study however suggests that further investigation of behavioral developmental effects of antiandrogens could be of future relevance. Analysis of the expression of genes related to neuronal growth, memory and other cognitive behaviors associated with behavioral changes, would then be of interest for further studies.
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Testes ecotoxicológicos para avaliação do potencial impacto ambiental em corpos receptores por efluente de Estação de Tratamento de Esgoto / Ecotoxicological tests to evaluate the potential environmental impact in receiving bodies by effluent from Sewage Treatment PlantIssa, Carina Graminha 22 March 2019 (has links)
O crescimento das cidades sem planejamento adequado tem como consequências a ampla geração e incorreta destinação de resíduos sólidos e líquidos e a falta de esgotamento sanitário adequado, resultando em impactos negativos ao meio ambiente e aos corpos hídricos. O esgoto doméstico é composto predominantemente por matéria orgânica, além de nitrogênio, fósforo, organismos patogênicos e elementos potencialmente tóxicos (e.g.: fármacos), que podem ou não ser removidos nas estações de tratamento de esgoto. Esse efluente final (esgoto tratado), é encaminhado para o corpo receptor, normalmente um rio ou córrego. Quando não removidos durante os processos de tratamento, os compostos remanescentes no esgoto podem causar alterações físicas, químicas e principalmente biológicas nos corpos receptores. Entre as principais alterações biológicas destaca-se a perda de biodiversidade aquática presente no corpo hídrico receptor. O presente trabalho avaliou o possível impacto ambiental causado pelo efluente final de Estação de Tratamento de Esgoto (E.T.E.) em corpos hídricos receptores por meio de testes ecotoxicológicos. Foram selecionadas três estações de tratamento de esgoto, as quais foram avaliadas. Foram coletadas amostras do afluente bruto (P0), efluente final (P1), um ponto a montante (P2) e um ponto a jusante (P3) da mistura do esgoto com o corpo receptor de cada estação. Foram realizados ensaios de toxicidade aguda utilizando as espécies de invertebrados aquáticos Daphnia magna, Chironomus sancticaroli, Allonais inaequalis. Os organismos foram expostos às amostras coletadas para avaliação da sobrevivência. Para a E.T.E. 1, foi constatado adequada eficiência de remoção de toxicidade do esgoto quando comparado o afluente bruto ao efluente tratado, e não foram observados impactos do lançamento do efluente no corpo receptor. A E.T.E. 2, mostrou baixa eficiência de remoção de toxicidade do afluente bruto para o efluente tratado, refletindo em potencial tóxico a longo prazo ao corpo receptor. Da mesma forma, a E.T.E. 3 se mostrou ineficiente para remoção de toxicidade do afluente bruto para o efluente tratado, que refletiu em potencial tóxico ao corpo receptor em longo prazo. De acordo com os resultados obtidos no presente estudo, ressalta-se a importância da utilização de testes ecotoxicológicos como complementação aos parâmetros físicos e químicos exigidos pela legislação para avaliação da eficiência de remoção de contaminantes presentes no esgoto e que possuem potencial para geração de impacto a curto ou longo prazo à biota aquática presente no corpo hídrico receptor. / The increase of cities, without adequate planning, results in the wide generation and incorrect destination of solid and liquid wastes and the lack of adequate sanitary structure, resulting in negative impacts on the environment. Domestic sewage is composed predominantly of organic matter, in addition to nitrogen, phosphorus, pathogenic organisms and potentially toxic elements (e.g., drugs), which may or may not be removed at sewage treatment plants. This final effluent (treated sewage) is conveyed to water bodies, usually a river or stream. When not removed during the treatment processes, the remaining compounds in the sewage can cause physical, chemical and especially biological changes in the water bodies. Among the main biological changes is the loss of aquatic biodiversity present in the receiving water body. Thus, the present study evaluated the possible environmental impact caused by the final effluent from Sewage Treatment Plant (S.T.P.) in receiving water bodies by means of ecotoxicological tests. Three sewage treatment stations were selected. Samples of the raw tributary (P0), final effluent (P1) and the upstream (P2) and downstream (P3) points of the sewage mixture with the receiving body of each station were collected. Acute tests were carried out using the aquatic species Daphnia magna, Chironomus sancticaroli, Allonais inaequalis. The organisms were exposed to the samples collected for evaluation of survival in acute tests. For E.T.E. 1, it was found good toxicity removal efficiency of the station when comparing the raw affluent to the treated effluent, and no impacts of effluent release on the receiving body were observed. E.T.E. 2, showed low toxicity removal efficiency from the raw affluent to the treated effluent, reflecting long term toxic potential to the water body. Likewise, E.T.E. 3 was shown to be inefficient to remove toxicity from the raw affluent to the treated effluent, which reflected toxic potential to the water body in long term. According to the results obtained in the present study, the importance of the use of ecotoxicological tests as complementation of the physical and chemical parameters required by the legislation to evaluate the efficiency of removal of pollutants present in the sewage that have potential to generate impact in the short or long term to aquatic biota present in the body water recipient.
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Geoestatística e Ecotoxicologia como ferramentas para gestão de reservatórios / Geostatistics and Ecotoxicology as tools for reservoir ecotoxicology management. Case study of Reservoir Guarapiranga (São Paulo, BR)Leal, Philipe Riskalla 14 February 2017 (has links)
Neste trabalho, objetivou-se o avaliar ecotoxicidade potencial do sedimento do Reservatório Guarapiranga da cidade de São Paulo (Brasil) frente a contaminação por metais, a fim de discutir sua qualidade e sua atual política de gerenciamento ambiental. Os metais analisados foram: Al, Cd, Cr, Cu, Fe, Mn, Pb e Zn. Utilizou-se das ferramentas de sistema de informação geográfica, geoprocessamento de imagens e da geoestatística para avaliação da dispersão espacial dos metais ao longo do sedimento do Reservatório, considerado por este trabalho como aquela região de maior implicação ecotoxicológica para contaminantes metálicos. Analisaram-se também as variáveis granulometria, matéria orgânica, nitrogênio total, fósforo total e umidade do sedimento, a fim de se estudar quimiometria do sedimento, a fim de melhor compreender as distribuições dos contaminantes ao longo do sedimento. Denominou-se esta classe de variáveis quimiométricas de parâmetros estruturais do sedimento. Observou-se grande variabilidade quantitativa e espacial entre os metais analisados, denotando que o Reservatório Guarapiranga sofre contaminação por diferentes vias no sistema. Observou-se falta de correlação quimiométrica entre os parâmetros estruturais do sedimento com os contaminantes metálicos, denotando grande variabilidade interna desse sistema. Referente a política de manejo do Reservatório, especialmente relativa ao controle algal e cianofítico, observou-se um gerenciamento de grande impacto ecossistêmico, focado mais no custo imediato do que em planejamentos de longo prazo. Tal gerenciamento, hoje, traz risco ecotoxicológico ao reservatório, mas também toxicidade potencial à própria população abastecida pelo reservatório quanto àquela sob exposição direta a este reservatório por meio de recreação, esportes náuticos, ou alimentação / In this work, the metal ecotoxicological potential of an urban reservoir\'s sediment has been assessed, in order to discuss its management policies and its implications to the society and to the ecosystem. The Reservoir Guarapiranga has been selected as the model environment for this study, once it possess a long history since its damming, several studies regarding its water and sediment conditions and an easy accessibility. The metals of greater implication to the Guarapiranga\'s cause were the Al, Cd, Cr, Cu, Fe, Mn, Pb and Zn, being selected as the main parameters for this study. Several scientific areas have been applied in this work. Among them, it is cited the Geographical Information System, geostatistical analysis, imagem geoprocessing, geochemical and chemometric analysis and, finally, the ecotoxicology. In order to better understand the relationship between the sediment and its contaminants, this study also assessed the granulometry, organic matter, total nitrogen, total phosphorous and the umidity of the sediment as explainable variables, translated as chemometric variables. My results showed great variability among the metals analized, denoting that each metal has its own path of contamination (contamination source point) in the reservoir, but a main direction (axis) of dispersion on the sediment. This later, mostly due to hydrological and eolic aspects of the ecosystem. Also, it\'s been observed lack of correlation between the chemometric and the metals parameters of the sediment, which also demonstrated the variation and stochasticity of the ecosystem. Lastly, the reservoir\'s environemntal management policies specially related to the algal and cyanophitic controlhave been analized through an ecotoxicological prisma, and it been denoted that it lacks long-term considerations, focusing only at punctual or short to middle-term solutions,resulting in the terrible ecotoxicological condition observed in the Guarapiranga\'s reservoir. Here, I punctuate the importance of new policy making, and its importance to the ecosystem and to the society, once hazardous condition of a reservoir is unberable not only through a ecosystemperspective, but as a sociological one
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Geoestatística e Ecotoxicologia como ferramentas para gestão de reservatórios / Geostatistics and Ecotoxicology as tools for reservoir ecotoxicology management. Case study of Reservoir Guarapiranga (São Paulo, BR)Philipe Riskalla Leal 14 February 2017 (has links)
Neste trabalho, objetivou-se o avaliar ecotoxicidade potencial do sedimento do Reservatório Guarapiranga da cidade de São Paulo (Brasil) frente a contaminação por metais, a fim de discutir sua qualidade e sua atual política de gerenciamento ambiental. Os metais analisados foram: Al, Cd, Cr, Cu, Fe, Mn, Pb e Zn. Utilizou-se das ferramentas de sistema de informação geográfica, geoprocessamento de imagens e da geoestatística para avaliação da dispersão espacial dos metais ao longo do sedimento do Reservatório, considerado por este trabalho como aquela região de maior implicação ecotoxicológica para contaminantes metálicos. Analisaram-se também as variáveis granulometria, matéria orgânica, nitrogênio total, fósforo total e umidade do sedimento, a fim de se estudar quimiometria do sedimento, a fim de melhor compreender as distribuições dos contaminantes ao longo do sedimento. Denominou-se esta classe de variáveis quimiométricas de parâmetros estruturais do sedimento. Observou-se grande variabilidade quantitativa e espacial entre os metais analisados, denotando que o Reservatório Guarapiranga sofre contaminação por diferentes vias no sistema. Observou-se falta de correlação quimiométrica entre os parâmetros estruturais do sedimento com os contaminantes metálicos, denotando grande variabilidade interna desse sistema. Referente a política de manejo do Reservatório, especialmente relativa ao controle algal e cianofítico, observou-se um gerenciamento de grande impacto ecossistêmico, focado mais no custo imediato do que em planejamentos de longo prazo. Tal gerenciamento, hoje, traz risco ecotoxicológico ao reservatório, mas também toxicidade potencial à própria população abastecida pelo reservatório quanto àquela sob exposição direta a este reservatório por meio de recreação, esportes náuticos, ou alimentação / In this work, the metal ecotoxicological potential of an urban reservoir\'s sediment has been assessed, in order to discuss its management policies and its implications to the society and to the ecosystem. The Reservoir Guarapiranga has been selected as the model environment for this study, once it possess a long history since its damming, several studies regarding its water and sediment conditions and an easy accessibility. The metals of greater implication to the Guarapiranga\'s cause were the Al, Cd, Cr, Cu, Fe, Mn, Pb and Zn, being selected as the main parameters for this study. Several scientific areas have been applied in this work. Among them, it is cited the Geographical Information System, geostatistical analysis, imagem geoprocessing, geochemical and chemometric analysis and, finally, the ecotoxicology. In order to better understand the relationship between the sediment and its contaminants, this study also assessed the granulometry, organic matter, total nitrogen, total phosphorous and the umidity of the sediment as explainable variables, translated as chemometric variables. My results showed great variability among the metals analized, denoting that each metal has its own path of contamination (contamination source point) in the reservoir, but a main direction (axis) of dispersion on the sediment. This later, mostly due to hydrological and eolic aspects of the ecosystem. Also, it\'s been observed lack of correlation between the chemometric and the metals parameters of the sediment, which also demonstrated the variation and stochasticity of the ecosystem. Lastly, the reservoir\'s environemntal management policies specially related to the algal and cyanophitic controlhave been analized through an ecotoxicological prisma, and it been denoted that it lacks long-term considerations, focusing only at punctual or short to middle-term solutions,resulting in the terrible ecotoxicological condition observed in the Guarapiranga\'s reservoir. Here, I punctuate the importance of new policy making, and its importance to the ecosystem and to the society, once hazardous condition of a reservoir is unberable not only through a ecosystemperspective, but as a sociological one
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