Impact of aquatic humic substances on speciation and toxicity of arsenic and cobalt to Ceriodaphnia dubia.

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.

Analysis of vitellogenin by histochemical method as an indicator of estrogenic effect in male Danio rerio exposed to metals.

Several chemicals present in the aquatic environment have the ability to alter the endocrine system of aquatic organisms, including the metals cadmium (Cd) and zinc (Zn). In males, a tool to analyze this effect is the vitellogenin (VTG) detection, a protein with defined function in the yolk production and subsequent aid to embryo-larval development. This protein is produced just by females and can be detected by simple and cheap methods such as histochemical method. Thus, this study aimed to analyze the capacity of Cd and Zn to induce VTG production in Danio rerio males and determine whether the histochemical labeling method is efficient to study estrogenic effects in this species. For this, D. rerio adult males were exposed chronically (21 days) to 0.25 and 1 µg/L of Cd and 120 and 180 µg/L of Zn, values allowed by the Brazilian and American legislation to aquatic life protection. After which, the organisms were submitted to a histological process as preparation to liver VTG marking by histochemical method and subsequent qualitative analysis of this protein in zebrafish's liver. After analyzes, it was possible observed that Cd and Zn are capable of inducing VTG production in D. rerio males and that the histochemistry method is efficient for detection of estrogenic effects in this species.

The Effects of Sediment Classification Pattern on a Water Column Organism, Ceriodaphnia dubia.

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.

Estudo da toxicidade de metais (zinco e cádmio) sobre Ceriodaphnia dubia, por multivias de exposição e recuperação biológica de descendentes / Study of metals toxicity (zinc and cadmium) to Ceriodaphnia dubia, for multi-exposition and biological recovery of offspring

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.

Toxicity assessment of arsenic and cobalt in the presence of aquatic humic substances of different molecular sizes.

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.

Remediation efficiency of three treatments on water polluted with endocrine disruptors: Assessment by means of in vitro techniques.

Chemical substances with potential to disrupt endocrine systems have been detected in aquatic environments worldwide, making necessary the investigation about water treatments able to inhibit such potential. The present work aimed to assess the efficiency for removing endocrine disruptors (with estrogenic and androgenic activity) of three simple and inexpensive substrates that could be potentially used in sectors or regions with limited resources: powdered activated carbon (PAC), powdered natural zeolite (ZEO) (both at a concentration of 500 mg L-1) and natural aquatic humic substances (AHS) (at 30 mg L-1). MilliQ-water and mature water from fish facilities (aquarium water, AW), were artificially spiked with 17ß-estradiol (E2), 17α-ethinylestradiol and dihydrotestosterone. Moreover, effluent samples from waste water treatment plants (WWTP) were also submitted to the remediation treatments. Estrogenic and androgenic activities were assessed with two cell lines permanently transfected with luciferase as reporter gene under the control of hormone receptors: AR-EcoScreen containing the human androgen receptor and HER-LUC transfected with the sea bass estrogen receptor. PAC was efficiently removing the estrogenic and androgenic compounds added to milliQ and AW. However, androgenic activity detected in WWTP effluents was only reduced after treatment with ZEO. The higher surface area of PAC could have facilitated the removal of spiked hormones in clean waters. However, it is possible that the substances responsible of the hormonal activity in WWTP have adsorbed to micro and nanoparticles present in suspension that would have been retained with higher efficiency by ZEO that show pores of several microns in size.

A comparative study of different diets to optimize cultivation of Hyalella azteca in the laboratory.

The aim of this study was to assess the influence of four diets on the laboratory cultivation of H. azteca, seeking to improve production of young specimens through reduced mortality and better growth and number of offspring per couple. The best diet was composed of a mixture of diluted commercial fish food, diluted yeast and primrose oil, associated with fish food flakes containing spirulina. With this diet the maximum mortality was 25 percent (at the end of 28 days), the average number of offspring/couple was 11.4 ± 2.8 (at the end of twelve days) and the peak weight and length of the males (at the end of 40 days) were 0.930 µg and 5.26 ± 1.1mm, respectively.

Analyses of the sediment toxicity of Monjolinho River, São Carlos, São Paulo State, Brazil, using survey, growth and gill morphology of two fish species (Danio rerio and Poecilia reticulata)

The objective of this work was to evaluate the toxicity of the sediments of the Monjolinho River (São Carlos - São Paulo/Brazil), through partial chronic toxicity bioassays with juveniles of D. rerio and P. reticulata. Histological analyses of the gills and biometric measurements were conducted to detect the possible deleterious effects that caused the death. In all points the sampled the alterations were found in the gills (hyperplasia, fusion of secondary lamellas and dilation of blood vessels), with the most intense lesions (second stage) occurring at the point two. The biometric analysis pointed to inadequate conditions for the growth of the test-organisms when exposed to the sediment samples. These results showed that the conditions of these environments were not suitable for the survival and growth of these fishes.

Este trabalho teve como objetivo avaliar a toxicidade dos sedimentos do rio Monjolinho (São Carlos - São Paulo/Brasil), por meio de bioensaios de toxicidade crônicos parciais com juvenis de D. rerio e P. reticulata. Foram realizadas análises histológicas das brânquias dos organismos para detectar possíveis efeitos deletérios que antecedam a mortalidade. Em todos os pontos amostrados, foram verificadas alterações nas brânquias (hiperplasia, fusão de lamelas secundárias e dilatação de vasos sanguíneos), sendo no ponto dois foram encontradas lesões mais acentuadas (segundo estágio). A análise biométrica dos indivíduos expostos em amostras de sedimento demonstrou condições inadequadas para o crescimento e desenvolvimentos dos organismos-teste em todo o ambiente estudado. Os sedimentos apresentaram maior toxicidade no período chuvoso, sendo estes resultados confirmados por meio das análises histológicas das brânquias dos organismos expostos, as quais demonstraram lesões mais acentuadas no mesmo período. Tal situação possivelmente decorre dos impactos antrópicos os quais o sistema está suscetível (efluentes domésticos, industriais e agrícolas), que foram constatados pelas análises físicas e químicas dos sedimentos.

The toxic potential of aldrin and heptachlor on Danio rerio juveniles (Cypriniformes, Cyprinidae).

With the objective of evaluating the effects of organochlorine pesticides (aldrin and heptachlor) on the survival, growth and gill morphology of juvenile zebrafish (Danio rerio), four partial chronic toxicity bioassays were conducted (seven days' duration) with both compounds in a semi-static system with renewal every 24 h. The results did not show any effects on the fish's survival, but did on their growth and gill morphology. Aldrin was more toxic than heptachlor, since the chronic value (CV) for growth was nearly 8.7 times less, a result confirmed by analyzing the histology of the gills, in which the changes detected were considered more severe for the former substance. In general, they were found branchial lesions of first stage, in other words, cell proliferation between the secondary lamellae, hyperplasia, lifting of respiratory epithelial cells; fusion of several secondary lamellae and dilation of blood vessels. Aneurysms (alterations of second stage) they were verified in the exposed organisms to the aldrin, which also presented about 10% of reduction in the standard length and 30% of reduction in the total weight in relation to the exposed organisms to the control. For heptachlor these values were of 8% and 25%, respectively. The intensity of the gill lesions and growth of the fish did not depend on the pesticide concentration, suggesting different modes of action of the products.

Alterations on growth and gill morphology of Danio rerio (Pisces, Ciprinidae) exposed to the toxic sediments

The objective of this work was to assess the toxicity of sediment samples from six cascade reservoirs in Tietê river system, in Säo Paulo State, Brazil, through chronic-partial toxicity bioassays with Danio rerio larvae as test-organisms. Histology of gills and biometric measurements were used to determine the existence of toxicity problems. The alterations on gill morphology here detected were considered of first stage (hyperplasia, lamellar junction and excess of mucous cells) in the first reservoirs and very slight in the last ones. The biometric analysis pointed to inadequate conditions for the growth of the test-organisms when exposed to the sediment of the rivers and upstream reservoirs and also indicated an improvement of environmental conditions along the system