Yeast as a model system to study genetic and post-translational regulation of metabolic pathways in mammals

Dissertation presented to obtain the Ph.D degree in Biology

Toxicity of carcinogenic and non-carcinogenic Polycyclic Aromatic Hydrocarbons and their mixtures to aquatic organisms under ecologically-relevant scenarios

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous priority pollutants that tend to be trapped in aquatic sediments due to their high hydrophobicity. Nonetheless, the differential toxicological effects and mechanisms between the various classes of PAHs and their mixtures, as they invariably occur in the environment, are scarcely known, especially under ecologically-relevant scenarios. This thesis aimed at establishing a bridge between the study of mechanistic pathways and environmental monitoring of carcinogenic and non-carcinogenic PAHs, by introducing ecological-relevance in the research with model PAHs. A first bioassay conducted in situ with the mussel Mytilus edulis demonstrated that, dredging operations in harbours increase PAH bioavailability, eliciting genotoxicity, and showed that established environmental guidelines underestimate risk. Subsequent ex situ bioassays were performed with the carcinogenic benzo[b]fluoranthene (B[b]F) and non-carcinogenic phenantrene (Phe), selected following preceding results, and revealed that low-moderate concentrations of these PAHs in spiked sediments induce genotoxic effects to the clam Ruditapes decussatus, therefore contradicting the general notion that bivalves are less sensitive to PAHs than vertebrates due to inefficient bioactivation. Also, it was demonstrated that passive samplers permit inferring on PAH bioavailability but not on bioaccumulation or toxic effects. On the other hand, sea basses (Dicentrarchus labrax), yielded a complex pattern of effects and responses, relatively to genotoxicity, oxidative stress and production of specific metabolites, especially when exposed to mixtures of the PAHs which led to additive, if not synergistic, effects. It was shown that Phe may elicit significant genotoxicity especially in presence of B[b]F, even though the low, albeit realistic, exposure concentrations diluted dose- and time-independent relationships. The present work demonstrated that environmental quality guidelines underestimate the effects of PAHs in realistic scenarios and showed that the significant genotoxic and histopathological effects caused by mixed PAHs may not be reflected by oxidative stress- or CYP-related biomarkers. Besides important findings on the metabolism of PAH mixtures, the work calls for the need to re-evaluate the criteria for assessing risk and for the disclosure of more efficient indicators of toxicological hazard.

Injuries and envenoming by aquatic animals in fishermen of Coxim and Corumbá municipalities, state of Mato Grosso do Sul, Brazil: identification of the causative agents, clinical aspects and first aid measures

INTRODUCTION: The fishes of continental Brazil have socioeconomic importance due to their potential for sport fishing and commercial and subsistence uses, as seen in the Upper Paraguay River Basin, particularly in the municipalities of the Pantanal region, where it is the second largest economic activity. Injuries caused in professional fishermen are common and poorly studied, as in other regions of the country. METHODS: Data were obtained from questionnaires and interviews with 100 professional fishermen, 50 in each municipality, between December 2008 and October 2009. RESULTS: All the fishermen reported some kind of injury caused by fish stings (78% of injuries) and fish, alligator and snake bites (22%) on the hands (46% of cases) and feet (35% of cases). Most of the patients had mild symptoms. The most severe cases were associated with secondary bacterial infections and required specific treatment and prolonged recovery associated with social and economic losses. CONCLUSIONS: The results of this study indicate that the stressful work conditions, inattention to basic preventive measures and carelessness were factors that contributed to accidents and that the toxicity and ability to inflict mechanical trauma of some aquatic species, plus the ineffective use of first aid and hospital treatment, contributed to the high morbidity and complications in many cases. Data from this study are relevant to the fishing communities of the Pantanal region, since they reveal high rates of accidents, lack of knowledge concerning first aid, initial treatment, injury prevention and lack of medical follow-up of the population.

Injuries caused by aquatic animals in Brazil: an analysis of the data present in the information system for notifiable diseases

AbstractINTRODUCTION:We present a review of injuries in humans caused by aquatic animals in Brazil using the Information System for Notifiable Diseases [ Sistema de Informação de Agravos de Notificação (SINAN)] database.METHODS:A descriptive and retrospective epidemiological study was conducted from 2007 to 2013.RESULTS:A total of 4,118 accidents were recorded. Of these accidents, 88.7% (3,651) were caused by venomous species, and 11.3% (467) were caused by poisonous, traumatic or unidentified aquatic animals. Most of the events were injuries by stingrays (69%) and jellyfish (13.1%). The North region was responsible for the majority of reports (66.2%), with a significant emphasis on accidents caused by freshwater stingrays (92.2% or 2,317 cases). In the South region, the region with the second highest number of records (15.7%), jellyfish caused the majority of accidents (83.7% or 452 cases). The Northeastern region, with 12.5% of the records, was notable because almost all accidents were caused by toadfish (95.6% or 174 cases).CONCLUSIONS:Although a comparison of different databases has not been performed, the data presented in this study, compared to local and regional surveys, raises the hypothesis of underreporting of accidents. As the SINAN is the official system for the notification of accidents by venomous animals in Brazil, it is imperative that its operation be reviewed and improved, given that effective measures to prevent accidents by venomous animals depend on a reliable database and the ability to accurately report the true conditions.

Observations on aquatic Macrophyte Dynamics In the Reservoir Of The Balbina Hydroelectric Powerplant, Amazonas State, Brazil

Between 1991 and 1995 aquatic macrophyte composition was observed in the lower part of the reservoir of the hydroelectric power plant of Balbina (Amazonas, Brazil). After closure of the dam in 1987, vegetation cover - mostly Eichhornia crassipes - was high, but was not quantified. After 1990 it declined rapidly with a characteristic succession pattern: Eichhornia ® Vincularia + Cyperaceae ® Salvinia. The Cyperaceae, and many other less dominant species, were mostly associated with drift wood, produced by the decomposing, emergent forest. Comparison of the chemical data of the Uatumã river before the construction of the dam (1983) with those of later years (1989 - 1995) suggests that the succession was the result of a relatively mild and short period of eutrophication, followed by declining nutrient levels. Annual variation of water levels, followed by aquatic and terrestrial decomposition of the marginal vegetation, may allow for the maintenance of relatively productive vegetation belts along the shore lines of islands and inundated stream valleys.

Aquatic and semiaquatic Heteroptera (Insecta) from Pitinga, Amazonas, Brazil

A list of aquatic and semiaquatic Heteroptera from a collecting trip to Pitinga, a village in a mining area in the County of Presidente Figueiredo in the Central Amazon is presented. Identified were fifty five species of Heteroptera, distributed in 13 families. Among the insects collected, some are new records for this Amazonian region and in addition 3 apparently undescribed species of Microvelia and one of Paravelia remain for further study.

Updated checklist of aquatic macrophytes from Northern Brazil

Field collection and herbaria data did not allow to quantify the diversity of aquatic plants from Northern Brazil, so we could not detect biogeographic patterns. Therefore, our objectives were to identify and quantify the aquatic macrophytes of North Brazilian states, analyzing herbaria data plataforms (SpeciesLink and Flora do Brasil). The checklist was produced by bibliographic search (articles published between 1980 and 2000), herbaria collections of the platforms SpeciesLink and Flora do Brasil and field expeditions, where we utilized asystematic sampling. We also analyzed the floristic similarity of aquatic macrophytes among Northern Brazil, wetlands of distinct Brazilian regions and the Neotropics. We recorded 539 species, of which 48 are endemic to Brazil. The states with highest number of species were Amazonas and Pará, independently on platform. The most represented families were Poaceae (89 species), Podostemaceae (55), Cyperaceae (50) and Fabaceae (47). We highlight the unprecedent richness of Podostemaceae, due to our own field collection efforts on favorable habitats, 25 species being endemic. Emergent and/or amphibious plants (515) were dominant in total species richness and were best represented in lotic habitats. We found significant differences in richness and floristics among states, obtained from the platforms. There is floristic similarity among Northern states and other Brazilian wetlands. In conclusion, we observed a rich aquatic flora in Northern Brazil, in spite of scarcity of records for Acre, Rondonia and Tocantins; we highlight the unprecedent number of endemic species of Podostemaceae (25) and contrasting richness between SpeciesLink and Flora do Brasil.

Trap efficiency evaluation for small mammals in the southern Amazon

The effectiveness of ecological researches on small mammals strongly depends on trapping techniques to survey communities and populations accurately. The main goal of this study was to assess the efficiency of three types of traps (Sherman, Tomahawk and Pitfall) to capture non-volant small mammals. We installed traps in 22 forest fragments in the southern Brazilian Amazonia. We captured 873 individuals belonging to 21 species; most of the individuals (N = 369) and species (N = 19) were trapped using Pitfalls, followed by Shermans (N = 271 individuals; N = 15 species) and Tomahawks (N = 233 individuals; N = 15 species). Pitfalls trapped a richer community subset of small mammals than the two other types of traps, and a more abundant community subset than Tomahawks. Proechimys sp. was the most abundant species trapped (N = 125) and Tomahawk was the most efficient type of trap to capture this species (N = 97 individuals). Neacomys spinosus and Marmosops bishopi were more trapped in Pitfalls (N = 92 and 100 individuals, respectively) than Shermans and Tomahawks. Monodelphis glirina was more trapped in Shermans and Pitfalls than Tomahawks. Species composition trapped using the three types of traps were distinct. Pitfalls captured a more distinct subset of the small mammal community than the two other live traps. We recommend the association of the three types of traps to reach a more comprehensive sampling of the community of small mammals. Thus, as stated by previous studies, we also recommend the complementary use of Shermans, Tomahawks and Pitfalls to account for a thorough sampling of the whole small mammal community in researches conducted in the tropical forests of Amazonia.

Zebrafish embryos as in vivo model for toxicity evaluation: screening of nanoparticle formulations DODAB:MO and DODAC:MO

Dissertação de mestrado em Molecular Genetics

Impacto de las prácticas agrícolas en los recursos hídricos de la provincia de Córdoba: aislamiento y caracterización de bacterias degradadoras de pesticidas para su utilización en procesos de biorremediación. Impact of agricultural activities on aquatic environments of Córdoba: isolation and characterization of bacteria with pesticide removal capacities for bioremediation treatments.

Los sistemas de agua dulce constituyen fuentes vitales para el desarrollo de la vida. Entre otras causas, el excesivo y poco controlado uso de pesticidas por las prácticas agrícolas modernas ha contribuido con la degradación de estos ecosistemas en la provincia de Córdoba (Secretaría de Ambiente, 2008). Los pesticidas son compuestos tóxicos y químicamente estables en el ambiente. Diversas especies microbianas presentan la capacidad para mineralizar dichos compuestos, siendo uno de los mecanismos de descomposición más importante. En el presente trabajo se pretende (1) Detectar y cuantificar principios activos de pesticidas en diferentes aguas ambientales de la región centro-sur de la provincia de Córdoba; (2) Aislar y caracterizar especies bacterianas a partir de muestras de aguas superficiales y subterráneas de la región que demuestren ser eficientes en la biodegradación de diferentes pesticidas. Para ello, se estudiará la presencia de 10 pesticidas organoclorados y organofosforados (lindano, 2,4D, DDT, p,p-DDE, �-endosulfán, �-endosulfán, clorpirifós, dimetoato) y herbicidas (atrazina) en muestras de agua superficiales y subterráneas (8 - 12m de profundidad) de la región agrícola centro-sur de Córdoba. Se establecerán diferentes puntos de muestreo en las principales cuencas hidrográficas: Río Tercero y Embalse de Río Tercero, Canal Desviador de Bell Ville, Laguna La Salada, Río Saladillo, Río Carcarañá y Río Cuarto. La determinación de pesticidas se realizará mediante Cromatografía de Gases (GC). Conjuntamente, se realizará el aislamiento de microorganismos a partir de las muestras de agua suplementadas con diferentes concentraciones de pesticidas (López et al., 2005) y se procederá a la caracterización morfológica y bioquímica (Lechevalier, 1989). Se realizarán curvas de crecimiento (DO600) y se determinará la viabilidad celular mediante el método de recuento en placa. El potencial catabólico de cada aislamiento se determinará analizando la concentración residual de pesticidas en el sobrenadante de los cultivos (Benimeli et al, 2003) y mediante ensayos de resting-cell (Hernández et al, 2008). Finalmente se realizará la caracterización genética (Weisburg, 1991) de los aislamientos que demuestren una mayor eficiencia en la biodegradación de pesticidas. El presente estudio pretende abordar una temática prioritaria como es la contaminación ambiental de ecosistemas acuáticos de la provincia de Córdoba. la detección de principios activos de pesticidas sería, por lo tanto, un indicador de contaminación de origen antropológica y brindaría información respecto al deterioro de la calidad del agua. Por otra parte, el aislamiento de bacterias adaptadas a las condiciones ecológicas de la región y capaces de metabolizar eficientemente diferentes pesticidas como fuentes de carbono y energía sería beneficioso para su utilización en futuros ensayos de biorremediación.