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A ação antimicrobiana da nanopartícula de prata (AgNP) favorece seu uso em diversos produtos. Sua toxicidade está relacionada com o tamanho da nanopartícula, seu estado de agregação e a capacidade em gerar espécies reativas de oxigênio (EsRO). O objetivo deste trabalho foi avaliar os efeitos citogenotóxicos, como também o estresse oxidativo em eritrócitos de Trachinotus carolinus expostos à AgNP. Os peixes receberam, via injeção intraperitoneal, três diferentes doses de suspensão de AgNP, 3, 1.5 e 0.75 μgAgNP/gpeixe. Após 12, 24, 36, 72 e 108 horas o sangue foi coletado para realizar o ensaio cometa e o teste do micronúcleo (MN) outras anormalidades nucleares (ANE), como também o fígado para a atividade enzimática da catalase. A AgNP demonstrou ser citogenotóxica, como também se capaz de promover o estresse oxidativo em Trachinotus carolinus. Os resultados mostram que os danos ao DNA e a ocorrência de MN e ANE apresentaram relação dose-resposta dependente. A redução no dano ao DNA mostrou estar relacionada com o aumento da atividade da catalase. Silver nanoparticles (AgNP) are applied as antimicrobial agents to many manufactured products. Nanoparticles size, aggregation and its induction of reactive oxygen species (ROS) are associated with AgNP toxicity. This study was undertaken to examine the citogenotoxicity as well as oxidative stress of AgNP in Trachinotus carolinos erythrocytes. The fish received tree different doses of 3, 1.5 e 0.75 µgAgNP/gfish by intraperitoneal injection. Bloods samples were collected and at 12, 24, 36, 72 and 108 hours post-injection to perform the comet assay and the micronucleus test. Extract of liver were used to measure catalase activity. This study showed that AgNP is citogenotoxic to this species and is able to induce oxidative stress. The results showed that the DNA damage, micronucleus and nuclear abnormalities was dose dependent. The reduction of DNA damage exhibit a close relationship with catalase activity.

Series of biomonitoring surveys were undertaken weekly in February 2012 to investigate the genotoxicity of the shallow waters around the Brazilian Antarctic Station "Comandante Ferraz" (EACF). The comet assay was applied to assess the damage to the DNA of hemocytes of the crustacean amphipods Gondogeneia antarctica collected from shallow waters near the Fuel Tanks (FT) and Sewage Treatment Outflow (STO) of the research station, and compare it to the DNA damage of animals from Punta Plaza (PPL) and Yellow Point (YP), natural sites far from the EACF defined as experimental controls. The damage to the DNA of hemocytes of G. antarctica was not significantly different between sites in the biomonitoring surveys I and II. In survey III, the damage to the DNA of animals captured in shallow waters near the Fuel Tanks (FT) and Sewage Treatment Outflow (STO) was significantly higher than that of the control site of Punta Plaza (PPL). In biomonitoring survey IV, a significant difference was detected only between the FT and PPL sites. Results demonstrated that the shallow waters in front of the station may be genotoxic and that the comet assay and hemocytes of G. antarctica are useful tools for assessing genotoxicity in biomonitoring studies of Antarctic marine coastal habitats. Séries de biomonitoramentos foram executadas semanalmente, durante o mês de fevereiro de 2012, para se investigar a genotoxicidade de ambientes costeiros rasos no entorno da Estação Antártica "Comandante Ferraz (EACF). O ensaio cometa foi aplicado para se avaliar os danos ao DNA hemocitário de crustáceos anfípodes Gondogeneia antarctica coletados em áreas rasas próximas aos tanques de armazenamento de combustível (FT) e saída de efluentes da estação de tratamento de esgoto (STO), em comparação aos danos ao DNA de animais provenientes de Punta Plaza (PPL) e Yellow Point (YP), locais naturais distantes da EACF, definidos como controles experimentais. O dano ao DNA hemocitário de G. antarctica não foi significativamente diferente entre locais, nos biomonitoramentos I e II. No biomonitoramento III, o dano ao DNA de animais coletados em águas rasas próximas aos tanques de combustível e saída de efluentes de esgoto foi significativamente maior do que aquele do controle de Punta Plaza. No biomonitoramento IV, a diferença foi significativa somente entre os locais dos tanques de combustível e de Punta Plaza. Estes resultados demonstram que a contaminação das águas em frente à EACF pode ser genotóxica e que, tanto o ensaio cometa quanto os hemócitos de G. antarcticasão ferramentas úteis na avaliação da genotoxicidade em estudos de biomonitoramento de habitats marinhos costeiros da Antártica.

Os hidrocarbonetos policíclicos aromáticos são tóxicos e/ou genotóxicos para diversos organismos marinhos e a toxicidade de muitos deles aumenta consideravelmente quando expostos à radiação ultravioleta artificial ou natural. O presente estudo visou avaliar a fototoxicidade do antraceno sobre a mortalidade e o dano ao DNA, segundo o ensaio cometa, de juvenis de pampos da espécie Trachinotus carolinus. Os peixes foram previamente expostos ao antraceno por 24 horas no escuro nas concentrações de 8, 16 e 32µg/L. e então transferidos para a água limpa e expostos à luz de lâmpadas fluorescentes ou à radiação ultravioleta B artificial (RUVB-35µW/cm2). Grupos controle (água e solvente) foram submetidos às mesmas condições experimentais. As exposições na ausência da radiação ultravioleta foram realizadas por períodos cumulativos de 10, 20 e 30h (5h/dia). Para as exposições à RUVB foram utilizados períodos desde 2 até 10h. Na ausência de radiação ultravioleta o antraceno não revelou ser tóxico, mas sua genotoxicidade, medida como danos ao DNA, apresenta resposta dose-dependente. A RUVB é letal para pampos. A exposição ao antraceno seguida de 2h de incidência de RUVB acelera a morte dos peixes em relação à mortalidade dos animais não contaminados. Este resultado revelou a fototoxicidade do antraceno para a espécie. O ensaio cometa pode ser considerado uma ferramenta sensível e útil para avaliar o efeito genotóxico de HPAs em peixes marinhos costeiros. Polycyclic aromatic hydrocarbons are toxic and/or genotoxic to a variety of marine organisms and under solar or artificial ultraviolet radiation acute toxicity of many of them can be substantially enhanced. The aim of the present study was to examine the effect of the phototoxicity of anthracene on the mortality and damage to the DNA, assessed by the comet assay, of juveniles of Florida pompanos, Trachinotus carolinus. The fish were pre-contaminated in the dark by three concentrations of 8, 16 and 32 µg/L of anthracene for 24 hours then transferred to clean seawater to be exposed to fluorescent light or to artificial ultraviolet B radiation (UVB - 35µW/cm2). Control groups (water and ethanol) were carried out in the same experimental conditions. The exposures to fluorescent light were carried out in cumulative periods of 10, 20 and 30 hours (5h/day). The exposures to UVB were conducted during periods of 2 to 10 hours. In the absence of ultraviolet radiation, the anthracene did not prove to be toxic, but its genotoxicity, evaluated as damage caused to the DNA, proved to be dose dependent. The UVB is lethal to Florida pompanos. The exposure of fish to the anthracene following UVB exposure for 2 hours resulted in earlier and higher mortality when compared to uncontaminated fish. These results indicated that anthracene is phototoxic to the pompanos. The comet assay can be considered a sensitive tool to evaluate the genotoxic effect of PAHs in coastal marine fish.

The comet assay was applied to the biomonitoring of genotoxicity in shallow waters around the Brazilian Antarctic Station “Comandante Ferraz” (EACF). Mean values of DNA damage to animals captured from shallow waters nearby the Fuel Tanks (FT) and Sewage Treatment Outflow (STO) were significantly higher than the values of the controls captured from shallow waters of Punta Plaza (PPL) and Yellow Point (YP), naturally undisturbed places far from the EACF.

Nanoparticles have physicochemical characteristics that make them useful in areas such as science, technology, medicine and in products of everyday use. Recently the manufacture and variety of these products has grown rapidly, raising concerns about their impact on human health and the environment. Adverse effects of exposure to nanoparticles have been reported for both terrestrial and aquatic organisms, but the toxic effects of the substances on marine organisms remain poorly understood. The main aim of this study was to evaluate the genotoxicity of TiO2-NP in the marine fish Trachinotus carolinus, through cytogenotoxic methods. The fish received two different doses of 1.5 μg and 3.0 μg-TiO2-NP g(-1) by intraperitoneal injection. Blood samples were collected to analyze erythrocyte viability using the Trypan Blue exclusion test, comet assay (pH>13), micronucleus (MN) and other erythrocyte nuclear abnormalities (ENA) 24, 48 and 72 h after injection. The possible cell uptake of TiO2-NP in fish injected with the higher dose was investigated after 72 h using transmission electron microscopy (TEM). The results showed that TiO2-NP is genotoxic and potentially cytotoxic for this species, causing DNA damage, inducing the formation of MN and other ENA, and decreasing erythrocyte viability. TEM examination revealed that cell uptake of TiO2-NP was mainly in the kidney, liver, gills and to a lesser degree in muscle. To the extent of the authors' knowledge, this is the first in vivo study of genotoxicity and other effects of TiO2-NP in a marine fish.

In the present study, the polycyclic aromatic hydrocarbon (PAH) genotoxicity was investigated in a one-step predator–prey relationship with the trophic-related marine species. Florida pompanos were fed for 5 and 10 days with pink shrimp post larvae previously exposed to benzo( a )pyrene (BaP) concentrations. Parent BaP body burden was measured in samples of Farfantepenaeus brasiliensis . BaP metabolites were determined in bile samples of Trachinotus carolinus and DNA damage was assessed through the comet and erythrocyte nuclear abnormalities (ENAs) assays in fish erythrocytes. BaP body burden increased significantly with the PAH concentration in pink shrimp PLs as well as the fish bile BaP metabolites. Both, comet and ENAs assays indicated significant increase on erythrocyte DNA damage of Florida pompanos fed with BaP-exposed pink shrimp on both feeding periods. The trophic route of BaP genotoxicity is discussed as well as the PAH biotransformation as the inducing mechanism for the DNA damages observed.

Seabob shrimp Xiphopenaeus kroyeri is a marine species that lives in shallow waters of coastal environments, often impacted by polycyclic aromatic hydrocarbons (PAH) pollution. In the present study, seabob shrimp were exposed for 96 h to benzo[a]pyrene (BaP) at the nominal concentrations of 100, 200, 400 and 800 microg-L −1 . Animals of the control groups were exposed either to clean water or to the BaP-carrier (DMSO). At the end of the exposures, muscle tissues were sampled for BaP uptake assessment and hepatopancreas and hemolymph for EROD enzyme activity and hemocytes DNA damage, respectively. EROD activity and DNA damage increased significantly as a function of BaP exposure concentrations. Significant correlations between BaP uptake and both EROD activity and DNA damage suggest that they can be used as suitable tools for integrated levels of study on the biomarkers of PAH exposure.

The photo-induced toxicity of anthracene was investigated as the mortality in Antarctic shallow water amphipod, Gondogeneia antarctica, at different concentrations of anthracene and different periods of exposure to natural sunlight and artificial UVA and UVB radiations. When exposed to natural sunlight, animals contaminated in the dark and placed in clean water or in anthracene solutions showed different degrees of mortality, dose–time dependent. Effects were even more evident when these animals were exposed to artificial UVA or UVB radiations. Depuration seemed to be a slow process. The effects of UV radiation and anthracene alone and the effects of the interactions of these two stressors implied that solar radiation is an important parameter that deserves consideration in the environmental assessment of polycyclic aromatic hydrocarbons in Antarctic coastal waters. G. antarctica proved to be a good bioindicator for the phototoxicity of anthracene in Antarctic shallow waters.

Made available in DSpace on 2019-09-12T16:25:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2018 Resumo A produção da aquicultura vem crescendo em todo o mundo e o cultivo de peixes marinhos no Brasil é ainda muito recente. Condições intensivas de aquicultura podem causar estresse fisiológico ao organismo do cultivo, o que pode ser avaliado por biomarcadores citogenotóxicos. O objetivo desse estudo foi avaliar o efeito genotóxico das condições de cultivo nos eritrócitos de juvenis de beijupirás Rachycentron canadum usando o ensaio cometa e o ensaio de micronúcleo e outras anormalidades nucleares. Juvenis de beijupirá foram cultivados por 13 semanas em um tanque indoor com circulação aberta de água e em um tanque-rede. O ensaio comera e o ensaio de anormalidades nucleares detectaram um maior dano ao DNA e uma maior frequência de anormalidades nucleares em eritrócitos de peixes cultivados no tanque indoor. Results showed that two methods are complementary. Além disso, beijupirás foram injetados com β-naftoflavona (BNF) nas concentrações de 2mgkg-1 e 10mgkg-1 em condições controladas de laboratório e mantidos por 7 dias em tanques separados para um melhor entendimento dos mecanismos de resposta dessa espécie a uma substância tóxica. O ensaio cometa não detectou nenhuma diferença significativa entre os peixes injetados com BNF e os da condição controle, enquanto que o ensaio das anormalidades nucleares apresentaram diferenças significativas entre peixes injetados com BNF e os do controle. Os danos identificados pelo ensaio cometa são quebras reparáveis na fita do DNA, enquanto que as anormalidades nucleares são permanentes. Possivelmente, o período de de manutenção dos peixes nos tanques após a injeção foi suficiente para limpar a BNF dos organismos e reparar as quebras na fita do DNA. Como os beijupirás parecem responder muito bem a compostos genotóxicos, o ensaio cometa e o ensaio de anormalidades nucleares podem ser ferramentas úteis para monitorar as condições de cultivo. Abstract Aquaculture production is continuously growing worldwide, and marine fish farming in Brazil is still in its infancy. Intensive farming conditions may cause physiological stress to the cultured organism, which can be evaluated by citogenotoxic biomarkers. The aim of this study was to assess the genotoxic effect of the rearing conditions in red blood cells of juvenile cobia Rachycentron canadum by using comet assay and micronucleus and other nuclear abnormalities assay. Juvenile cobia were reared for 13 weeks in indoor tank with open water circulation and in near shore cage. The comet assay and the nuclear abnormalities assay detected higher DNA damage and higher nuclear abnormalities frequency in erythrocytes of fish reared in the indoor tank. Results showed that two methods are complementary. Additionally, cobia were injected with ß-naphthoflavone (BNF) at concentrations of 2mgkg-1 and 10mgkg-1 in laboratory controlled conditions, and maintained for 7 days in separate tanks to better understand the response mechanisms of this species to a toxic substance. The comet assay did not detect any significant differences between BNF injected and control fish, whereas nuclear abnormalities assay showed significant differences between BNF injected and the control groups. The damages identified by the comet assay are repairable breaks in the DNA strands, whereas nuclear abnormalities may be permanent. Possibly the period of maintenance after injection was enough to clean BNF from the organisms and to repair the breaks in the DNA strands. As cobia seems to respond very well to genotoxic elements, comet assay and nuclear abnormalities assay would be useful tools to monitor farming conditions. [Botelho, Marina Tenório; Passos, Maria José de Arruda Campos Rocha; Hasue, Fabio Matsu; Lemos, Daniel; Yasumaru, Fanny; Phan, Ngan van; Gomes, Vicente] Universidade de São Paulo, Brazil Rodrigues, Edson] Universidade de Taubaté, Brazil