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Humic substances (HS) interact with trace metals such as As and Co, affecting their mobility and availability in aquatic systems. However, their combined effects on toxicity to aquatic organisms are not totally understood. The objective of this study was to evaluate the toxicity of Co(II) and As(III) to the water flea Ceriodaphnia dubia in the presence of HS, considering element speciation. Toxicity assays were performed in the presence and absence of HS at two different concentrations of As(III) (10 and 20 µg/L) and Co(II) (50 and 100 µg/L). The free As(III) and Co(II) (< 1 kDa, fraction most potentially bioavailable) in the test solutions were determined via ultrafiltration. While free Co(II) decreased by approximately 80% in the presence of HS, free As(III) decreased just by 1%. Despite the higher percentage of As(III) potentially bioavailable, the presence of HS reduced significantly the toxicity of As at 20 µg/L (no toxicity was observed at 10 µg/L). This was attributed to direct effects of HS such as hormesis, hormone-like effects of HS and/or formation of protective coating. These effects also stimulated the reproduction, including in the assays in the absence of As and Co. HS reduced the toxicity of Co(II) at both test concentrations. The results of this investigation support that HS should be considered when safe limits for As and Co are defined.

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The sediment compartment stands out because it functions as both a temporary sink of pollutants and a potential source of these elements that may become available to the water column.This study aimed to correlate the concentrations of total metals in the crude sediment and in the interstitial water with the ecotoxicity in the water column using an a modified sediment ecotoxicity test with Ceriodaphnia dubia. The results indicate that the sediment may contribute to the toxicity in the water column and that such toxicity is possibly not related to the metals present. Based on the chemical analysis of the metals, the Canadian Sediment Quality Guidelines (SQGs) would frame the sediment as non-toxic to benthic organisms, but the SQGs have no reference standards for possible effects on nektonic organisms. Due to the complexity of this compartment, it is fundamental to evaluate the interactions of the different pollutants in the system and possible effects on the nektonic organisms.

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RESUMO Os metais frequentemente são avaliados em águas doces como soluções dissolvidas, assumindo que o efeito tóxico é causado unicamente por via aquática (respiração e contato). No entanto, estudos abrangendo concomitantemente a toxicidade na água e no alimento, como acontece no meio, são pouco discutidos na literatura. No presente estudo, a toxicidade de zinco e cádmio foi avaliada expondo-se Ceriodaphnia dubia simultaneamente ao alimento e ao meio aquoso. A espécie de alga verde Raphidocelis subcapitata foi exposta durante 96h a concentrações de Zn (0,18 e 0,27 mg.L-1) e Cd (0,001 e 0,0015 mg.L-1). Os resultados foram analisados estatisticamente por meio da Análise de Variância (Kruskal-Wallis). As algas foram usadas como fonte de alimento para C. dubia, durante exposição crônica (oito dias), nas mesmas concentrações. Posteriormente, os neonatos (geração F1) foram introduzidos em água e alimentação sem contaminantes, para averiguação da capacidade de recuperação biológica. Foram avaliados número de neonatos por indivíduos, morfologia dos neonatos e quantificação dos metais em tecido biológico. Os resultados demonstraram que nas concentrações testadas não houve inibição no crescimento de R. subcapitata, enquanto para C. dubia evidenciou-se toxicidade crônica pela redução na taxa reprodutiva nas duas gerações, para ambos metais. Concluiu-se que, mesmo em concentrações relativamente baixas, os metais zinco e cádmio podem alterar o padrão reprodutivo dos invertebrados de água doce, comprometendo o ecossistema aquático e sua capacidade de recuperação. Considerando os efeitos tóxicos desses metais e sua interferência no sistema biológico, novos ensaios ecotoxicológicos devem ser realizados para melhor compreensão do comportamento dessas substâncias nos organismos.

ABSTRACT Metals are often evaluated in fresh water as dissolved solutions, assuming that the toxic effect is caused only by water (respiration and contact). However, toxicity studies in food and water, in a concomitant way, as occurs in the environment, are less discussed. In this study, zinc and cadmium toxicity was evaluated through the exposure of Ceriodaphnia dubia to contaminated food and water. The species of green algae Raphidocelis subcapitata was exposed for 96h to concentrations of Zn (0,18 and 0,27 mg.L-1) and Cd (0,001 and 0,0015 mg.L-1). The results were statistically analyzed by means of Analysis of Variance (Kruskal-Wallis). Algae were used as food source for C. dubia, during chronic exposure (eight days) in the same concentrations described. Subsequently, neonates (F1 generation) were introduced in non-contaminated water and food in order to ascertain their biological recovery capacity. The number of newborns by individuals, morphology of newborns, and quantification of metals in biological tissue were evaluated. The results showed that the tested concentrations did not inhibit the growth of R. subcapitata, while C. dubia had chronic toxicity, with reduction in the reproductive rate in both generations, for both metals. The results allowed the conclusion that, even at relatively low concentrations, the metals zinc and cadmium can alter the reproductive patterns of freshwater invertebrates, compromising the aquatic ecosystem and its resilience. Thus, considering the toxic effects of these metals and their interference in the biological system, new ecotoxicological tests should be performed to allow a better understanding of the behavior of these substances in organisms.

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The release of contaminants in aquatic ecosystems can be influenced by humic acids. In this study, toxicity tests using environmentally relevant concentrations of arsenic and cobalt were conducted both in the presence and absence of aquatic humic substances (AHS) and the fractions of different molecular sizes in the range of (<5, 5-10;10-30; 30-100 and >100kDa) using the microcrustacean Ceriodaphnia dubia. AHS together with arsenic reduced the toxicity, and the toxicity decreased in fractions of larger molecular size AHS. Despite the presence of cobalt, the reduction in toxicity was not observed and that depended on the molecular size of AHS. There was a trend of enhanced toxicity for Co in fractions of larger molecular sizes, opposed to that found for arsenic. Thus, the humic substances alter toxicity of trace elements, and this effect varies depending on the size of the humic substances.

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