Mechanisms of immunotoxic effects of nanomaterial in fish

Detalhes bibliográficos
Autor(a) principal: Costa, Leonor Coutinho
Data de Publicação: 2013
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10773/12616
Resumo: Information and knowledge related to nanotechnology raise new challenges to the scientific community mainly in terms of human health risks and environmental implications associated to nanomaterials. In this perspective, the contamination of aquatic environments cannot be overlooked since it is an ultimate repository of the contaminants where this emerging appears as part of a cocktail of different classes of contaminants. Thus, the major task of this work was to connect gaps in current knowledge with a comprehensive sequence of biological responses toward environmentally relevant concentrations of engineered nanomaterials (IONM - silica coated iron oxide nanomaterial functionalized with dithiocarbamate group) and their interaction with other conventional anthropogenic contaminant (Hg - mercury), outlining the interaction with the innate immune system of fish. The research was divided into following steps: i) phagocytes macrophages were isolated from peritoneum (P-phagocytes), gill (G-phagocytes), head kidney (HK-phagocytes) and spleen (S-phagocytes) of European eel Anguilla anguilla L. in order to evaluate whether, and how can IONM and its co-exposure to Hg modulate phagocytes status and function; ii) determine the changes in phagocytes activation and their association to peroxidative damage (OBA - oxidative burst activity; LPO - lipid peroxidation); iii) to assess the impact of IONM on phagocytes enzymatic (CAT - catalase; GPX - glutathione peroxidase; GR - glutathione reductase; and GST - glutathione S-transferase) and non-enzymatic (NP-SH - non-protein thiols; and TGSH - total glutathione) antioxidants protection overtime. It was hypothesized that IONM can cause measurable changes in fish immune response and oxidative stress modulation. A period of exposure-dependency was exhibited by IONM alone and IONM+Hg joint exposures accrued impacts on A. anguilla phagocytes. IONM exposure alone lead to an acute response in terms of viability increase in P-phagocytes and modulated phagocytic activity in P-, and S-.phagocytes during 2 hours of exposure; whereas, IONM lead to a chronic immunotoxicity during 72 hours exposure only in S-phagocytes. However, IONM+Hg exposure lead to both acute and chronic response in terms of modulated phagocytic activity with no change in viability in P-, HK- and S-phagocytes only. Increase in the period of exposure to Hg disrupted phagocytic activity of P-, HK- and S-phagocytes, an increase in P- and decrease in HK- and S- phagocytes was perceptible at late hours of exposure. The occurrence of synergism between IONM and Hg was evidenced at 72 hours by significantly increasing trends of phagocytosis increase. A differential extent of OBA and LPO induction at the end of different period of exposure to IONM, Hg or IONM+Hg was also perceptible. The OBA induction and its concomitant association to LPO induction were observed only in gill after exposure to Hg (8 and 48 hours) and IONM+Hg (8 hours). At late hours of exposure, an induction was observed in G- phagocytes (OBA) after exposure to IONM+Hg suggesting that the concomitant exposure was unable to mitigate the Hg-accrued negative impacts. A. anguilla also displayed that the damage was accompanied with a differential modulation of enzymatic and non-enzymatic antioxidants in P, G-, HK- and S-phagocytes. Under IONM alone exposure, no LPO induction along the time was observed probably due to efficient induction of, GR and GST providing a better protection to IONM exposed phagocytes. However, antioxidants protection responses displayed hours of exposure dependency in G- and S-phagocytes where, an insufficiency of elevated CAT, GPX, GST, NP-SH and TGSH was clearly depicted for the maintenance of pro- and antioxidant balance optimum for scavenging ROS and protecting membrane lipids against IONM impact. As increased LPO was observed under IONM alone exposure condition, the joint action of IONM+Hg led to elevated damages to membrane-lipids at 4 and 8 hours (G-phagocytes), 2 hours (HK-phagocytes) and 24 hours (S-phagocytes) of exposure. These responses together, point towards the antioxidants defense failure for the protection of membrane lipids during those periods of exposure to IONM+Hg. However, it is important to underline here that during the late hours of exposure (72 hours), the results imply the positive effect of the concomitant exposure (IONM+Hg) which significantly mitigated the said negative impacts of Hg. Overall, the observations of this study open up new insight into the areas of evaluation of immune defense mechanisms in fish exposed to IONM and recommend that the interactions between IONM and other conventional anthropogenic contaminants should be considered while interpreting the fish immunotoxicity responses to IONM exposure in a multi-pollution state.
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spelling Mechanisms of immunotoxic effects of nanomaterial in fishQuímica analíticaToxicologia ambiental - Ecossistemas aquáticosNanomateriaisMercúrio - ToxicidadeInformation and knowledge related to nanotechnology raise new challenges to the scientific community mainly in terms of human health risks and environmental implications associated to nanomaterials. In this perspective, the contamination of aquatic environments cannot be overlooked since it is an ultimate repository of the contaminants where this emerging appears as part of a cocktail of different classes of contaminants. Thus, the major task of this work was to connect gaps in current knowledge with a comprehensive sequence of biological responses toward environmentally relevant concentrations of engineered nanomaterials (IONM - silica coated iron oxide nanomaterial functionalized with dithiocarbamate group) and their interaction with other conventional anthropogenic contaminant (Hg - mercury), outlining the interaction with the innate immune system of fish. The research was divided into following steps: i) phagocytes macrophages were isolated from peritoneum (P-phagocytes), gill (G-phagocytes), head kidney (HK-phagocytes) and spleen (S-phagocytes) of European eel Anguilla anguilla L. in order to evaluate whether, and how can IONM and its co-exposure to Hg modulate phagocytes status and function; ii) determine the changes in phagocytes activation and their association to peroxidative damage (OBA - oxidative burst activity; LPO - lipid peroxidation); iii) to assess the impact of IONM on phagocytes enzymatic (CAT - catalase; GPX - glutathione peroxidase; GR - glutathione reductase; and GST - glutathione S-transferase) and non-enzymatic (NP-SH - non-protein thiols; and TGSH - total glutathione) antioxidants protection overtime. It was hypothesized that IONM can cause measurable changes in fish immune response and oxidative stress modulation. A period of exposure-dependency was exhibited by IONM alone and IONM+Hg joint exposures accrued impacts on A. anguilla phagocytes. IONM exposure alone lead to an acute response in terms of viability increase in P-phagocytes and modulated phagocytic activity in P-, and S-.phagocytes during 2 hours of exposure; whereas, IONM lead to a chronic immunotoxicity during 72 hours exposure only in S-phagocytes. However, IONM+Hg exposure lead to both acute and chronic response in terms of modulated phagocytic activity with no change in viability in P-, HK- and S-phagocytes only. Increase in the period of exposure to Hg disrupted phagocytic activity of P-, HK- and S-phagocytes, an increase in P- and decrease in HK- and S- phagocytes was perceptible at late hours of exposure. The occurrence of synergism between IONM and Hg was evidenced at 72 hours by significantly increasing trends of phagocytosis increase. A differential extent of OBA and LPO induction at the end of different period of exposure to IONM, Hg or IONM+Hg was also perceptible. The OBA induction and its concomitant association to LPO induction were observed only in gill after exposure to Hg (8 and 48 hours) and IONM+Hg (8 hours). At late hours of exposure, an induction was observed in G- phagocytes (OBA) after exposure to IONM+Hg suggesting that the concomitant exposure was unable to mitigate the Hg-accrued negative impacts. A. anguilla also displayed that the damage was accompanied with a differential modulation of enzymatic and non-enzymatic antioxidants in P, G-, HK- and S-phagocytes. Under IONM alone exposure, no LPO induction along the time was observed probably due to efficient induction of, GR and GST providing a better protection to IONM exposed phagocytes. However, antioxidants protection responses displayed hours of exposure dependency in G- and S-phagocytes where, an insufficiency of elevated CAT, GPX, GST, NP-SH and TGSH was clearly depicted for the maintenance of pro- and antioxidant balance optimum for scavenging ROS and protecting membrane lipids against IONM impact. As increased LPO was observed under IONM alone exposure condition, the joint action of IONM+Hg led to elevated damages to membrane-lipids at 4 and 8 hours (G-phagocytes), 2 hours (HK-phagocytes) and 24 hours (S-phagocytes) of exposure. These responses together, point towards the antioxidants defense failure for the protection of membrane lipids during those periods of exposure to IONM+Hg. However, it is important to underline here that during the late hours of exposure (72 hours), the results imply the positive effect of the concomitant exposure (IONM+Hg) which significantly mitigated the said negative impacts of Hg. Overall, the observations of this study open up new insight into the areas of evaluation of immune defense mechanisms in fish exposed to IONM and recommend that the interactions between IONM and other conventional anthropogenic contaminants should be considered while interpreting the fish immunotoxicity responses to IONM exposure in a multi-pollution state.Informações e conhecimentos relacionados com a nanotecnologia colocam novos desafios à comunidade científica, principalmente em termos de riscos para a saúde humana e alterações ambientais. A contaminação dos ambientes aquáticos não pode ser ignorada, uma vez que é um repositório final dos contaminantes onde estes aparecem como parte de um conjunto complexo de diferentes tipos. O objetivo principal deste trabalho foi relacionar lacunas no conhecimento atual com uma sequência de respostas biológicas, para concentrações ambientalmente relevantes de nanomateriais (ONM - nanomateriais de óxidos de ferro revestidos com sílica e funcionalizados com grupos de ditiocarbamato) e avaliar a sua interação com outros contaminantes antropogénicos, nomeadamente mercúrio (Hg), na interação com o sistema imunitário de peixes. A investigação desenvolvida foi dividida nos seguintes passos: i) os macrófagos fagocitados foram isolados da cavidade peritoneal (P-fagócitos), das guelras (G-fagócitos), da “cabeça do rim” (HK-fagócitos) e do baço (S-fagócitos) da enguia europeia Anguilla anguilla L., a fim de avaliar como podem os ONM e a sua co-exposição com o mercúrio modular o estado das fagocitoses e a sua função; ii) avaliar as alterações na ativação das fagocitoses e a sua associação ao dano peroxidativo (OBA – atividade respiratória oxidativa; LPO - peroxidação lipídica); III) avaliar o impacto das ONMs ao longo do tempo nos antioxidantes, nomeadamente fagocioses enzimáticas (CAT - catalase; GPX - glutationa peroxidase; GR - glutationa redutase; e GST - Glutationa S-transferase) e não enzimáticas (NP-SH- proteína não-tiol; e TGSH - glutationa total). É apresentada a hipótese de que as ONMs podem causar mudanças na resposta imunológica de peixe e modular o stress oxidativo. Uma dependência do período de exposição foi observada para as ONMs isoladas e também uma sucessão de impactos sobre as fagocitoses da Anguilla Anguilla. A exposição isolada às ONMs parece poder induzir uma resposta aguda em termos de aumento de viabilidade em P-fagócitos e uma atividade fagocítica modulada em P e S-fagócitos após 2 horas de exposição; as ONMs podem induzir imunotoxicidade crónica durante uma exposição de 72 horas, apenas em S-fagócitos. A coexposição a ONM+Hg induziu uma resposta aguda e crónica em termos de atividade fagocítica modulada, com nenhuma mudança na viabilidade em P, HK e S-fagócitos. O aumento do período de exposição a Hg interrompeu a atividade fagocítica de P-, HK - e S-fagócitos. Porém, um aumento de P - e diminuição de HK - e S-fagócitos foi percetível nas 72 horas. A ocorrência de sinergismo entre as ONMs e o Hg foi evidenciado às 72 horas pela tendência do aumento significativo das fagocitoses. Diferença nas induções de OBA e LPO para diferentes períodos de exposição, às ONMs, Hg e ONMs+Hg foi também percetível. Uma indução de OBA paralela à resposta do LPO foi observada unicamente nas guelras por exposição ao Hg (8 e 48 horas) e ONMs+Hg (8 horas). Para períodos mais longos de exposição, foi observada uma indução nas G-fagócitos (OBA) depois da exposição a ONMs+Hg, sugerindo que a contaminação ONMs+Hg é capaz de mitigar os impactos negativos do Hg-acumulado ao longo do tempo. A. Anguilla evidenciou danos ao nível da modulação diferencial de antioxidantes enzimáticos e não enzimáticos em P, G, HK e S-fagócitos. Sob exposição simples às ONMs, não foi observada nenhuma indução do LPO ao longo do tempo, talvez devido à indução eficiente de GR e GST, proporcionando uma melhor proteção para os fagócitos expostos às ONMs. No entanto, as respostas de proteção dos antioxidantes exibiram dependência das horas de exposição em G - e S-fagócitos onde uma insuficiência elevada em CAT, GPX, GST, NP-SH e TGSH foi claramente observada para a manutenção do equilíbrio antioxidante e eliminação de ROS, protegendo os lipídios da membrana contra o impacto das ONMs. A ação conjunta das ONM+Hg conduziu a elevados danos nos lipídios da membrana às 4 e 8 horas (G-fagócitos), 2 horas (HK-fagócitos) e 24 horas (SP-fagócitos) de exposição. Estas respostas conjuntas, apontam para o fracasso da defesa dos antioxidantes na proteção dos lipídios da membrana durante os períodos de exposição com ONM+Hg. De salientar que para exposições de 72 horas, os resultados evidenciam efeitos positivos da exposição concomitante a ONM+Hg o que atenuou impactos negativos do Hg. As observações deste estudo dão novas ideias sobre a avaliação dos mecanismos de defesa imunotoxicológica em peixes expostos a ONMs e indicam que as interações entre ONMs e outros contaminantes antropogénicos devem ser consideradas na interpretação das respostas imunotóxicas de peixe expostos às ONMs em situações contaminação múltipla.Universidade de Aveiro2014-09-05T12:20:21Z2013-01-01T00:00:00Z2013info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10773/12616TID:201582392engCosta, Leonor Coutinhoinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-02-22T11:23:01Zoai:ria.ua.pt:10773/12616Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T02:48:45.309391Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Mechanisms of immunotoxic effects of nanomaterial in fish
title Mechanisms of immunotoxic effects of nanomaterial in fish
spellingShingle Mechanisms of immunotoxic effects of nanomaterial in fish
Costa, Leonor Coutinho
Química analítica
Toxicologia ambiental - Ecossistemas aquáticos
Nanomateriais
Mercúrio - Toxicidade
title_short Mechanisms of immunotoxic effects of nanomaterial in fish
title_full Mechanisms of immunotoxic effects of nanomaterial in fish
title_fullStr Mechanisms of immunotoxic effects of nanomaterial in fish
title_full_unstemmed Mechanisms of immunotoxic effects of nanomaterial in fish
title_sort Mechanisms of immunotoxic effects of nanomaterial in fish
author Costa, Leonor Coutinho
author_facet Costa, Leonor Coutinho
author_role author
dc.contributor.author.fl_str_mv Costa, Leonor Coutinho
dc.subject.por.fl_str_mv Química analítica
Toxicologia ambiental - Ecossistemas aquáticos
Nanomateriais
Mercúrio - Toxicidade
topic Química analítica
Toxicologia ambiental - Ecossistemas aquáticos
Nanomateriais
Mercúrio - Toxicidade
description Information and knowledge related to nanotechnology raise new challenges to the scientific community mainly in terms of human health risks and environmental implications associated to nanomaterials. In this perspective, the contamination of aquatic environments cannot be overlooked since it is an ultimate repository of the contaminants where this emerging appears as part of a cocktail of different classes of contaminants. Thus, the major task of this work was to connect gaps in current knowledge with a comprehensive sequence of biological responses toward environmentally relevant concentrations of engineered nanomaterials (IONM - silica coated iron oxide nanomaterial functionalized with dithiocarbamate group) and their interaction with other conventional anthropogenic contaminant (Hg - mercury), outlining the interaction with the innate immune system of fish. The research was divided into following steps: i) phagocytes macrophages were isolated from peritoneum (P-phagocytes), gill (G-phagocytes), head kidney (HK-phagocytes) and spleen (S-phagocytes) of European eel Anguilla anguilla L. in order to evaluate whether, and how can IONM and its co-exposure to Hg modulate phagocytes status and function; ii) determine the changes in phagocytes activation and their association to peroxidative damage (OBA - oxidative burst activity; LPO - lipid peroxidation); iii) to assess the impact of IONM on phagocytes enzymatic (CAT - catalase; GPX - glutathione peroxidase; GR - glutathione reductase; and GST - glutathione S-transferase) and non-enzymatic (NP-SH - non-protein thiols; and TGSH - total glutathione) antioxidants protection overtime. It was hypothesized that IONM can cause measurable changes in fish immune response and oxidative stress modulation. A period of exposure-dependency was exhibited by IONM alone and IONM+Hg joint exposures accrued impacts on A. anguilla phagocytes. IONM exposure alone lead to an acute response in terms of viability increase in P-phagocytes and modulated phagocytic activity in P-, and S-.phagocytes during 2 hours of exposure; whereas, IONM lead to a chronic immunotoxicity during 72 hours exposure only in S-phagocytes. However, IONM+Hg exposure lead to both acute and chronic response in terms of modulated phagocytic activity with no change in viability in P-, HK- and S-phagocytes only. Increase in the period of exposure to Hg disrupted phagocytic activity of P-, HK- and S-phagocytes, an increase in P- and decrease in HK- and S- phagocytes was perceptible at late hours of exposure. The occurrence of synergism between IONM and Hg was evidenced at 72 hours by significantly increasing trends of phagocytosis increase. A differential extent of OBA and LPO induction at the end of different period of exposure to IONM, Hg or IONM+Hg was also perceptible. The OBA induction and its concomitant association to LPO induction were observed only in gill after exposure to Hg (8 and 48 hours) and IONM+Hg (8 hours). At late hours of exposure, an induction was observed in G- phagocytes (OBA) after exposure to IONM+Hg suggesting that the concomitant exposure was unable to mitigate the Hg-accrued negative impacts. A. anguilla also displayed that the damage was accompanied with a differential modulation of enzymatic and non-enzymatic antioxidants in P, G-, HK- and S-phagocytes. Under IONM alone exposure, no LPO induction along the time was observed probably due to efficient induction of, GR and GST providing a better protection to IONM exposed phagocytes. However, antioxidants protection responses displayed hours of exposure dependency in G- and S-phagocytes where, an insufficiency of elevated CAT, GPX, GST, NP-SH and TGSH was clearly depicted for the maintenance of pro- and antioxidant balance optimum for scavenging ROS and protecting membrane lipids against IONM impact. As increased LPO was observed under IONM alone exposure condition, the joint action of IONM+Hg led to elevated damages to membrane-lipids at 4 and 8 hours (G-phagocytes), 2 hours (HK-phagocytes) and 24 hours (S-phagocytes) of exposure. These responses together, point towards the antioxidants defense failure for the protection of membrane lipids during those periods of exposure to IONM+Hg. However, it is important to underline here that during the late hours of exposure (72 hours), the results imply the positive effect of the concomitant exposure (IONM+Hg) which significantly mitigated the said negative impacts of Hg. Overall, the observations of this study open up new insight into the areas of evaluation of immune defense mechanisms in fish exposed to IONM and recommend that the interactions between IONM and other conventional anthropogenic contaminants should be considered while interpreting the fish immunotoxicity responses to IONM exposure in a multi-pollution state.
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