Proteomic approach to oxidative stress in Daphnia magna

The keystone aquatic organism Daphnia magna is extensively used to assess the toxicity of chemicals. This has recently lead to an increase in the omics literature focusing on daphnids, an increase fuelled by the sequencing of the Daphnia pulex genome. Yet, no omics study has looked directly at oxidative stress (OS) in daphnids, even though OS is of primary importance in the response of aquatic organisms to their changing environment and is often induced by anthropogenic xenobiotics. This thesis thus focuses on the application of redox-proteomics, the study of the oxidative modification of proteins, to D. magna Specifically, daphnids were exposed to copper or paraquat, two well studied prooxidants, and protein carbonyls were labelled with fluorescein-5-thiosemicarbazide prior to twodimensional electrophoresis (2DE). This showed clearly that both compounds affect a different portion of the proteome. The identified proteins indicated that energy metabolism was affected by paraquat, while copper induced a reduction of the heat shock response (heat shock proteins, proteases and chaperones) a counterintuitive result which may be adaptative to metal toxicity in arthropods. The same approach was then applied to the study of the toxicity mechanism of silver nanoparticles (AgNP), an increasingly utilised form of silver with expected environmental toxicity, and its comparison to silver nitrate. The results demonstrate that, although less toxic than silver ions, AgNP toxicity functions through a different mechanism. AgNP toxicity is thus not a product of silver dissolution and increased protein carbonylation indicates that AgNP cause OS. Interestingly three of the four tested compounds altered vitellogenin levels and oxidation. Vitellogenins could thus represent an interesting subproteome for the detection of stress in daphnids. Finally, an experiment with oxidised BSA demonstrates the applicability of solid phase hydrazide in the enrichment of undigested carbonylated proteins.

Nueva carta agroclimática de horas de frío en la Argentina

p.219-225

Multiple origins of European populations of giant liver fluke Fascioloides magna (Trematoda: Fasciolidae), an invasive liver parasite of ruminants

The giant liver fluke, Fascioloides magna, liver parasite of free-living and domestic ruminants of Europe and North America, was analysed in order to determine the origin of European populations and to reveal the biogeography of this originally North American parasite on the European continent. The previously selected variable fragments of the mitochondrial cytochrome c oxidase subunit I (cox1; 384 bp) and nicotinamid dehydrogenase subunit I (nad1; 405 bp) were applied as a tool. The phylogenetic trees and haplotype networks were constructed and the level of genetic structuring was evaluated using population genetic tools. In F. magna individuals originating from all European natural foci (Italy, Czech Republic, Danube floodplain forests) and from four of five major North American enzootic areas, 16 cox1 and 18 nad1 haplotypes were determined. The concatenated sequence set produced 22 distinct haplotypes. The European fluke populations were less diverse than those from North America in that they contained proportionately fewer haplotypes (8), while more substantial level of genetic diversity and higher number of haplotypes (15) were recorded in North America. Only one haplotype was shared between the European (Italy) and North American (USA/Oregon and Canada/Alberta) flukes supporting a western North American origin of the Italian F. magna population. Haplotypes found in Italy were distinct from those determined in the remaining European localities what indicates that introduction of F. magna onto the European continent is a result of more than one event. In Czech focus, a south-eastern US origin of giant liver fluke was revealed. Identical haplotypes, common for parasites from Czech Republic and from expanding focus of Danube floodplain forests, implies introduction of F. magna to the Danube region from an already established Czech focus.

Carta educacional do Concelho de Aljustrel como elemento impulsionador do enriquecimento educativo local

Efetuamos, numa primeira fase do trabalho, uma abordagem teórica, definindo alguns conceitos fundamentais para o desenvolvimento e fundamentação deste estudo. Ainda no campo teórico, realizámos a caraterização do território em estudo nas suas diversas vertentes. Esta investigação procura conhecer as instituições concelhias mais relevantes, caraterizar a sua atividade e perceber quais são os seus contextos de aprendizagem. Por parte das instituições, há disponibilidade para estabelecer parcerias e colaborar com a escola, podendo esta articulação contribuir para aumentar e diversificar a sua capacidade e oferta educativa, através da mobilização dos conhecimentos existentes nas instituições locais. O estudo empírico foi quantitativo, recorrendo ao inquérito por questionário como meio de recolha de dados. Foram estudadas 25 instituições num universo de 73 associações existentes. Procedeu-se à apresentação, análise e interpretação dos dados e principais conclusões do estudo. Foram feitas algumas recomendações e sugestões; ### ABSTRACT: We established a theorethical approach in the initial stage of our work, defining some crucial concepts for the development and grounding of this study. We also characterized the territory under study in its several forms. The research aims at knowing the most important municipal institutions, characterizing their activities and understanding what are their learning contexts. By the institutions, there is will to establish partnerships and co-operate with the school. This linkage may contribute to increase and diversify their ability and educational provision by harnessing existing knowledge in local institutions. The study was quantitative, using the inquiry and survey as a way to collect data. 25 institutions were studied within a total number of 72 existing associations. Afterwards, the data and main conclusions of the study were presented, analysed and interpreted. Some recommendations and suggestions were made.

Bisphenol A migration from plastic materials: direct insight of ecotoxicity in Daphnia magna

Bisphenol A (BPA) is an endocrine disrupting chemical (EDC) whose migration from food packaging is recognized worldwide. However, the real overall food contamination and related consequences are yet largely unknown. Among humans, children’s exposure to BPA has been emphasized because of the immaturity of their biological systems. The main aim of this study was to assess the reproductive impact of BPA leached from commercially available plastic containers used or related to child nutrition, performing ecotoxicological tests using the biomonitoring species Daphnia magna. Acute and chronic tests, as well as single and multigenerational tests were done. Migration of BPA from several baby bottles and other plastic containers evaluated by GC-MS indicated that a broader range of foodstuff may be contaminated when packed in plastics. Ecotoxicological test results performed using defined concentrations of BPA were in agreement with literature, although a precocious maturity of daphnids was detected at 3.0 mg/L. Curiously, an increased reproductive output (neonates per female) was observed when daphnids were bred in the polycarbonate (PC) containers (145.1±4.3 % to 264.7±3.8 %), both in single as in multigenerational tests, in comparison with the negative control group (100.3±1.6 %). A strong correlated dose-dependent ecotoxicological effect was observed, providing evidence that BPA leached from plastic food packaging materials act as functional estrogen in vivo at very low concentrations. In contrast, neonate production by daphnids cultured in polypropylene and non-PC bottles was slightly but not significantly enhanced (92.5±2.0 % to 118.8±1.8 %). Multigenerational tests also revealed magnification of the adverse effects, not only on fecundity but also on mortality, which represents a worrying trend for organisms that are chronically exposed to xenoestrogens for many generations. Two plausible explanations for the observed results could be given: a non-monotonic dose–response relationship or a mixture toxicity effect.

Produção e facilitação da visualização on-line de uma carta de desporto de natureza

Os Sistemas de Informação Geográfica detêm um papel de crescente importância para a tomada de decisão ao nível do ordenamento do território, conforme o que a própria legislação evidencia. Existe uma carência de uniformização da informação para que possa existir um cruzamento eficiente e uma disponibilização mais facilitada à população e às diversas entidades. As instituições públicas, como é o caso do Instituto de Conservação e Florestas (ICNF I.P.), reconhecem esta necessidade, pelo que se propôs uma série de tarefas, para a concretização deste estágio, no sentido de a colmatar. Este instituto detém um papel de especial relevo na conservação do património natural assim como no ordenamento das áreas protegidas para que se possa usufruir das mesmas, sob um ponto de vista cada vez mais ecológico e promotor de desenvolvimento sustentável. A elaboração de um instrumento de gestão territorial, como se pode classificar a Carta de Desporto de Natureza (CDN), acarreta grande responsabilidade e exige um forte empenho, não apenas no envolvimento da população e das entidades que dele fruirão, mas no cumprimento da legislação existente quer para as atividades quer para o próprio território, no sentido da sua proteção, preservação e promoção de usos adequados. Neste trabalho procurou-se cumprir todas estas disposições sempre com o objetivo de, ao se construir normas de uniformização e ao se elaborar um exemplo de CDN para um território com tanto valor como é a Reserva Natural do Estuário do Tejo, tornar acessível a informação sobre esta área protegida. A intensão é também promover a motivação para a prática de um desporto natureza, saudável e em equilíbrio com o ambiente. O desafio não se ficou por aqui, havendo também a procura de utilização de Free Open Source Software, para que o processo de utilização e disponibilização informação SIG seja cada vez mais acessível e direta.

Aplicación de un sistema de información geográfica y de la carta electrónica isoparalitoral en las evaluaciones hidroacústicas de la biomasa de recursos pesqueros en el mar peruano

Se describe la utilización de un Sistema de Información Geográfica (Software Mapinfo 4.0), para realizar cálculos de biomasa de recursos pesqueros en base a la obtención de datos de densidad acústica estratificados por áreas isoparalitorales. Para ello se digitalizó la carta isoparalitoral, en la que en el pasado se ploteaban manualmente los valores ecointegrados; el producto obtenido, una carta electrónica isoparalitoral, permite realizar con mayor precisión y rapidez los cálculos para obtener la biomasa de recursos pesqueros. En el documento se describe, a modo de ejemplo, la obtención de la biomasa de la anchoveta Engraulis ringens a propósito de la ejecución del Crucero 9811-12 de Evaluación Hidroacústica de Recursos Pelágicos llevado a cabo a bordo del BIC José Olaya Balandra entre los días 28 de noviembre y 23 de diciembre de 1998.

Ingria, e la Carelia Russa : tratte da una carta del Golfo di Finlandia publicata dal Banco di Agrimensura di Stockolm nel 1788. Ab. Borghi disegno.

Huomautus: Pasguali scrisse.

Palladii Rutilii Tauri Aemiliani de re rustica libri tredecim ; desiderantur octavi finis, nonus, decimus et undecimi pars magna.

Puteanus

Carta compromiso servicio acompañamiento para arranque de la empresa

Desarrollo empresarial y creación de empresa

Behavioural, mensural, and ecological correlates of polygyny in the eastern meadowlark (Sturnella magna)

During 1982 and 1983 I studied male attributes and attributes of the territory of male Eastern Meadowlarks (Sturnella magna) in order to determine whether there was a correlation between any of the attributes investigated and the number of females attracted by a male. Seventeen males, nine of which were polygynous and eight monogamous, were studied in 1982 and sixteen males.of which .. seven were polygynous and nine ~onogamous, were studied in 1983. The study was conducted in Short Hills Park, 10 km southwest of St. Catharines, Ontario and was designed to compare two hypotheses: the "sexy son" hypothesis (Weatherhead and Robertson,1977) and the polygyny threshold model (Verner and Willson,1966, Orians, 1969). Male attributes investigated were male size and song behaviour. Six measures of male size were taken: weight, flattened and natural wing chord length, culmen length, bill depth and length of the tarsometatarsus. In 1983 song repertoire size and song versatility measures were investigated. Attributes of the territory studied were: territory size, density of plant stems, percentage plant cover and measures of vegetation structure. In 1983 Arthropods were collected from each territory and sorted according to taxonomic group and size. During 1983, territory attributes were sampled twice, once early and once later in the nesting season. Analysis of data involved univariate comparisons between monogamous and polygynous males using T-tests and multivariate comparisons were made using discriminant function analysis (DFA) and principle components analysis (PCA).No correlations were found between the number of females attracted with, .ny measure of male size or with me, .sures of song versatili or size of song repertoire. Also no correlation was found between terri size and the number of females nesting on a terri . Some attributes of the male's terri id distinguish between monog,mous and po s males of thistudy. Analysis of Arthropod numbers showed that e~ .eran counts were significantly great~r on polygynous territories, a1 the total numb~rs of Arthropods collected showed no s fico .nt differences between territories of monogamous and po males. DFA chose ear teran and Hymenopteran counts as multivariate discriminators; both variables we' e more vegetation revealed that there were no univariate differences between the two groups of males fOT 1982 stem densities, but ~ spp. and Solidago spp. were chosen DFA as multivariate discriminators. The total number of plant stems and of Vicia spp. stems were s ficantly the early 1983 ing on monogamous territories for however DFA found no multivariate discriminators" Variables concerned with the overall aspects of vegetation structure showed significant differences between territories of monogamous and polygynous males. DFA of the 1982 sampling of vegetation structure showed significantly greater mat depth and vegetation height on polygynous territories, a finding which was not supported, however, by peA. For the early 1983 sampling period, plant height was greater on polygynous territories. Multivariate analysis identified greater green cover on polygynous territories, greater ground cover on monogamous territories, and greater depth of mat material on monogamous territories as discriminators between territories of monogamous and polygynous males. A DFA on the major variables of the study showed no significant difference between the territories of monogamous and polygynous male Meadowlarks. Of the correlations found, some were for non-prey Arthr~ods, for cover plants with very small samples sizes, or for variables which were greater for monogamous males during one sampling period and polygynous males during the next. While multivariate discriminators were found, peA showed no grouping of monogamous or polygynous males according to any of the variables investigated. On the basis of the univariate and multivariate analysis of major variables, I concluded that there were no correlations between the number of females attracted with male attributes and no unambiguous correlation with attributes of the territory. My study does not unequivocally support either the "sexy son" or the polygyny threshold hypothesis.