Adrislaine da Silva Mansano

Acute and chronic toxicity of diuron and carbofuran to the neotropical cladoceran Ceriodaphnia silvestrii

In order to contribute to the increase of the body of knowledge on the sensitivity of tropical indigenous species to pesticides, acute and chronic toxicity tests were conducted with the neotropical cladoceran Ceriodaphnia silvestrii. Tests were carried out with the active ingredients diuron and carbofuran and one of their commercial formulations, the Diuron Nortox® 500 SC and the Furadan® 350 SC, respectively. For carbofuran, the active ingredient was more toxic than the commercial product, whereas for diuron, the commercial product appeared more toxic. In addition, hormetic effects on fertility were recorded for intermediate diuron concentrations. Acute and chronic toxicity data indicated that C. silvestrii was among the most sensitive invertebrate species for both test compounds. Based on concentrations measured in Brazilian water bodies, these compounds represent ecological risks for causing direct and indirect toxic effects on C. silvestrii and other aquatic organisms. Our results support previous claims on the advantages of using native species to better tune ecological risk assessment of chemicals in tropical ecosystems.

The use of rotifers as test species in the aquatic effect assessment of pesticides in the tropics

The present study aimed at evaluating the suitability of rotifers as standard invertebrate test species for the aquatic effect assessments of pesticides, with special emphasis to tropical settings. This was done by weighing rotifers against the criteria that are traditionally used for this end. Rotifers are easy to maintain and culture in the laboratory and their (biological) response to chemical stressors like pesticides is well known. As abundant organisms in aquatic ecosystems, they play a key role in energy flow and nutrient cycling. Although they are often considered to have a low sensitivity to pesticides, a sensitivity analysis conducted in this study revealed that they may be more sensitive than the standard invertebrate test species Daphnia magna to fungicides. In addition, few toxicity data were available for rotifers other than Brachionus calyciflorus and these data were almost exclusively acute (EC50) toxicity values. Subsequently, the sensitivity of other rotifers as well as the chronic sensitivity, bioaccumulation potential, and possible role in biomagnification of pesticides in aquatic foodwebs remains largely unknown. Given their greater diversity and ecological role in tropical freshwaters as compared to temperate freshwaters, the use of rotifers in tropical risk assessments and immediate research needs are discussed.

Toxicity of abamectin and difenoconazole mixtures to a Neotropical cladoceran after simulated run-off and spray drift exposure

Aquatic risk assessments of pesticides in tropical countries have often been disputed for being largely based on risk evaluations conducted in temperate regions. Although pesticide sensitivity comparisons between temperate and tropical freshwater organisms have indeed not revealed consistent differences, risk assessments are currently still based on a relatively small tropical toxicity dataset. In addition, greater levels of runoff and spray drift may be expected in tropical than in temperate agroecosystems, indicating that aquatic life in edge-of-field water bodies is likely to be subjected to higher concentrations of pesticides and their mixtures. The aim of the present study was to evaluate the toxicity of Kraft® 36 EC (a.i. abamectin), Score® 250 EC (a.i. difenoconazole) and their mixture to the Neotropical cladoceran Macrothrix flabelligera. Laboratory toxicity tests with the individual formulated products indicated EC50-48h values of 3.1 and 659μg a.i./L given as nominal test concentrations, respectively. Mixtures of the two pesticides revealed a concentration-dependent deviation of the independent action model, with antagonism at low and synergism at high pesticide mixture concentrations. Laboratory toxicity tests were also conducted with microcosm water that was treated with the individual or mixtures through runoff or direct overspray. Microcosm tanks receiving runoff water from experimental soil plots applied with recommended doses of the individual pesticides did not show toxicity to the test organism. Microcosms that received runoff water containing the pesticide mixture, however, did cause a short-term effect on immobility. The microcosms that were treated by direct overspray of both pesticide formulations showed the most pronounced toxic effects. Study findings suggest a potential risk of these pesticides at environmentally relevant concentrations, especially when they are both present.

Effects of diuron and carbofuran and their mixtures on the microalgae Raphidocelis subcapitata

In aquatic environments, organisms are often exposed to mixtures of several pesticides. In this study, the effects of carbofuran and diuron and their mixtures on the microalgae Raphidocelis subcapitata were investigated. For this purpose, toxicity tests were performed with the single compounds (active ingredients and commercial formulations) and their combinations (only active ingredients). According to the results, the toxicity of active ingredients and their commercial formulations to R. subcapitata was similar. In the single exposures, both carbofuran and diuron inhibited significantly the R. subcapitata growth and caused physiological (chlorophyll a content) and morphological (complexity and cell size) changes in cells, as captured by flow cytometry single-cell properties. Regarding the mixture toxicity tests, data fitted to both reference models, concentration addition (CA) and independent action (IA), and evidenced significant deviations. After the CA fitting, dose-ratio dependent deviation had the best fit to the data, demonstrating synergism caused mainly by diuron and antagonism caused mainly by carbofuran. After fitting the IA model, a synergistic deviation represented the best fit for the diuron and carbofuran mixtures. In general, the two reference models indicated the occurrence of synergism in the mixtures of these compounds, especially when diuron was the dominant chemical in the combinations. The increased toxicity caused by the mixture of these pesticides could pose a greater environmental risk for phytoplankton. Thus, exposure to diuron and carbofuran mixtures must also be considered in risk assessments, since the combination of these compounds may result in more severe effects on algae population growth than single exposures.

Life cycle traits of Philodina roseola Ehrenberg, 1830 (Rotifera, Bdelloidea), a model organism for bioassays

This paper describes experimental results on the life cycle of the rotifer Philodina roseola cultured in the laboratory. Detailed information on life-cycle parameters of a certain species provides a deep understanding and contributes to a better knowledge of the role of the species in the community, besides providing data that are basic to other ecological investigations such as secondary production estimates and knowledge for applications such as its utilization as test-organism in ecotoxicological studies. The average duration of embryonic development of P. roseola was 23.88 h, the age at maturity of primipara was 3.5 days and the maximum lifespan was 23 days. The average size of the rotifer neonate was 198.77 µm, the mean size of primipara was 395.56 µm and for adults 429.96 µm. The average fecundity was 1.22 eggs per female per day and the mean number of eggs produced per female during the entire life was 22.33. The deceleration of somatic growth from the start of the reproductive stage represents a trade-off between growth and reproduction that is often seen in micrometazoans. The life history of P. roseola follows the strategy of other bdelloid species characterized by a rapid pre-reproductive development and canalization of most assimilated energy to reproduction after reaching maturity. The differences observed in total fecundity and longevity between our P. roseola cultures and those from previous studies were probably due to differences of intrinsic adaptation of this species ecotypes to the conditions of their natural environments.

Seasonal variation of the protozooplanktonic community in a tropical oligotrophic environment (Ilha Solteira reservoir, Brazil)

The seasonal variation of the protozooplanktonic community (ciliates and testate amoebae) was studied in a tropical oligotrophic reservoir in Brazil, which was under the influence of two contrasting climatic seasons (rainy/warm and dry/cold). The aim of this study was to evaluate the effect of these climatic changes on physical, chemical and biological variables in the dynamic of this community. The highest mean density of total protozoans occurred in the rainy/warm season (5683.2 ind L-1), while the lowest was in the dry/cold (2016.0 ind L-1). Considering the seasonal variations, the protozoan groups that are truly planktonic, such as the oligotrichs (Spirotrichea), predominated in the dry season, whereas during the rainy season, due to the material input and resuspension of sediment, sessile protozoans of the Peritrichia group were the most important ones. The dominant protozoans were Urotricha globosa, Cothurnia annulata, Pseudodifflugia sp. and Halteria grandinella. The highest densities of H. grandinella were associated with more oxygenated and transparent water conditions, while the highest densities of C. annulata occurred in sites with high turbidity, pH and trophic state index (TSI). The study demonstrated that density and composition of protozooplanktonic species and groups of the reservoir suffered seasonal variation due to the environmental variables (mainly temperature, turbidity, water transparency, dissolved oxygen and TSI) and the biological variables (e.g. morphological characteristics, eating habits and escape strategies from predation of the species).

Protozooplankton and its relationship with environmental conditions in 13 water bodies of the Mogi-Guaçu basin - SP, Brazil

Protozooplankton is an important component of the aquatic microbial food webs and its composition, density, and distribution reflect the chemical, physical, and biological aspects of the environment. Considering the scarce literature on freshwater protozoans in Brazil and on protozoan ecology in subtropical environments, we listed the ciliates and amoebae taxa found in 13 water bodies in Sao Paulo State and analyzed their abundance in relation to the environmental variables. We collected two samples in each environment, fixed immediately with mercuric chloride and stained with bromophenol blue. After microscopical analysis, 74 protozoan genera were identified and the Ciliophora were dominant in the majority of the environments. The Stichotrichia, represented mostly by the genus Halteria, occurred in all environments, and was the dominant subclass in five of them. The canonic correspondence analysis of the most frequent genera and the environmental variables showed that nitrite and nitrate were the variables that better explained the distribution of Limnostrombidium, Urotricha, and Vorticella. The densities of the genera Halteria, Coleps, and of the species Cinetochilum margaritaceum were positively affected by increasing concentrations of dissolved oxygen, particulate phosphate, conductivity, and temperature. C. margaritaceum were also negatively affected by increasing concentrations of nitrite and nitrate. Considering that we made only one sampling in each environment, the richness was high compared to the mean diversity of lakes in the Sao Paulo State. The Diogo Lake, located in an ecological reserve, was the richest one, confirming the need of more research on the biodiversity of more preserved environments.

Sensitivities of three tropical indigenous freshwater invertebrates to single and mixture exposures of diuron and carbofuran and their commercial formulations

As compared to their temperate counterparts, few toxicity tests have been conducted so far into the evaluation of the sensitivity of indigenous tropical species to pesticides. Especially mixture toxicity assessments appear to be scarce. To contribute to increase our knowledge in this arena, we evaluated the acute toxicity of diuron and carbofuran and their mixtures to the neotropical oligochaetes Allonais inaequalis and Dero furcatus, and the ostracod Strandesia trispinosa. Tests were performed with both the pure active ingredients, as well as their formulated products. The toxicity of the latter to the three test organisms was generally greater than that of the pure active ingredients, although absolute differences were rather small. The sensitivity of the indigenous species was slightly greater than temperate test species from the same taxonomic groups. The concentration addition conceptual model best described the results of the mixture toxicity data. Derived deviations of this model appeared to be dependent on the test organism and as to whether the pesticides were applied as active ingredients or their commercial products. Reported field concentrations of the two pesticides indicate risks to freshwater biota, especially if they are both present. The test species used in the present study are concluded to be suitable candidates as surrogate test organisms in local pesticide risk evaluations.

Protozoans bacterivory in a subtropical environment during a dry/cold and a rainy/warm season

In aquatic ecosystems, bacteria are controlled by several organisms in the food chain, such as protozoa, that use them as food source. This study aimed to quantify the ingestion and clearance rates of bacteria by ciliates and heterotrophic nanoflagellates (HNF) in a subtropical freshwater reservoir (Monjolinho reservoir - São Carlos - Brazil) during one year period, in order to verify their importance as consumers and controllers of bacteria in two seasons, a dry/cold and a rainy/warm one. For this purpose, in situ bacterivory experiments were carried out bimonthly using fluorescently labeled bacteria with 5-(4,6 diclorotriazin-2yl) aminofluorescein (DTAF). Although ciliates have shown the highest individual ingestion and clearance rates, bacterivory was dominated by HNF, who showed higher population ingestion rates (mean of 9,140 bacteria h−1 mL−1) when compared to ciliates (mean of 492 bacteria h−1 mL−1). The greater predation impact on bacterial communities was caused mainly by the small HNF (< 5 μm) population, especially in the rainy season, probably due to the abundances of these organisms, the precipitation, trophic index state and water temperature that were higher in this period. Thus, the protozoan densities together with environmental variables were extremely relevant in determining the seasonal pattern of bacterivory in Monjolinho reservoir.

TOXICIDADE AGUDA DO AGROTÓXICO CARBOFURANO AO CLADÓCERO CERIODAPHNIA SILVESTRII DADAY, 1902

RESUMO: Nos ultimos anos o aumento do consumo de agrotoxicos em todo o mundo tem preocupado agencias nacionais e internacionais em decorrencia dos diversos impactos ambientais e das serias implicacoes a saude humana. Dentre os agrotoxicos, o carbofurano e utilizado amplamente no Brasil em culturas de batata, milho, amendoim, arroz e soja. Como existe o risco deste composto ser carreado por diversos mecanismos ate os corpos d’agua e causar toxicidade a especies nao-alvo, e necessaria a avaliacao dos seus efeitos nestes ecossistemas. O objetivo deste trabalho foi avaliar a toxicidade aguda do carbofurano sobre a especie nativa tropical Ceriodaphnia silvestrii . Para isso, realizaram-se testes de toxicidade aguda, com concentracoes nominais deste agrotoxico. Para o dafinideo C. silvestrii , os valores de CE 50 24 h e CE 50 48 h foram de 1,80 µg L -1 e 0,75 µg L -1 , respectivamente. Os resultados obtidos mostraram que C. silvestrii apresentou elevada sensibilidade ao agrotoxico carbofurano e foi mais sensivel do que as especies de cladoceros de regioes temperadas, de acordo com os dados da literatura. Considerando-se estes resultados e da ampla distribuicao de C. silvestrii em regioes tropicais e subtropicais, sugere-se o uso desta especie em avaliacoes ecotoxicologicas com agrotoxicos em aguas doces tropicais. Palavras-chave: Microcrustaceo. Pesticida. Toxicidade.