Made available in DSpace on 2014-08-20T14:30:17Z (GMT). No. of bitstreams: 1 Dissertacao_Maria_Raquel_Fernandez_Morinigo.pdf: 1321282 bytes, checksum: 91f2eb8e65e4b67c642c6df035df822a (MD5) Previous issue date: 2009-04-28 / Aim: To evaluate the cytotoxicity and genotoxicity of sodium percarbonate in comparison with bleaching agents used in discolored pulpless teeth from a mouse fibroblast cell culture. Methodology: The bleaching agents cytotoxicity and genotoxity were evaluated both in their pure form of products as well as in concentration commonly used in clinical practice. Hydrogen peroxide (HP), carbamide peroxide (CP), sodium perborate (SP) and sodium percarbonate (SPC) were diluted in DMEM in series. To evaluate the cytotoxicity, the survival of 3T3 mouse fibroblasts were measured photometrically by using MTT assay after a 24h exposure period. Genotoxicity was indicated by the micronuclei (MN) formation and the modification of the normal cell was analyzed by light microscopy (400x). The statistical analysis was performed by one-way ANOVA, followed by a multiplecomparison Tukey post hoc test (p < 0.05). Results: All tested groups exhibited a dose-dependent cytotoxicity. However, CP has showed a similar cytotoxic effect when compared with DMEM untreated control (UC) group. PH and SPC were significantly more cytotoxic than SP. Genotoxicity test has showed that SPC and SP have presented an intermediate rate of MN frequency over the UC group. The mean rate of MN frequency to HP was higher and statistically more significant than other groups tested. No difference was observed when CP and UC groups were compared. Conclusions: The results suggest that the cytotoxicity of SPC was similar to HP and significantly more cytotoxic than SP and CP. On the other hand, SPC and SP has shown an intermediate genotoxic effect, lower than HP and higher than CP, which was non-genotoxic / Objetivo: Avaliar a citoxicidade e genotoxidade do percarbonato de sódio e comparar com os agentes clareadores comumente utilizados no tratamento de dentes despolpados em cultura celular. Metodologia: A citotoxidade e genotoxicidade dos agentes clareadores foram avaliadas tanto na forma pura de cada produto quanto nas concentrações utilizadas na prática clínica. Peróxido de Hidrogênio (PH), peróxido de carbamida (PC), perborato de sódio (PS) e percarbonato de sódio (PCS) foram diluídos em DMEM. A viabilidade de fibroblastos da linhagem 3T3/NIH após 24h de exposição aos diferentes agentes foi medida fotometricamente por meio do teste colorimétrico com MTT. A genotoxicidade foi indicada pela formação e contagem de micronúcleos (MN) analisados em microscópio óptico comum (400x). A análise estatística dos dados foi feita utilizando o teste ANOVA de uma via complementado com o teste de comparações múltiplas Tukey post hoc (p < 0, 005). Resultados: Todos os grupos testados apresentaram um efeito citotóxico dosedependente. Entretanto, o PC mostrou citotoxicidade semelhante ao grupo controle (GC). PH e PCS foram significativamente mais citotóxicos que PS. O teste de genotoxicidade mostrou que PCS e PS apresentaram uma frequência intermediária de MN quando comparados ao GC. Por outro lado, a frequência média de MN usando PH foi significantemente superior aos outros grupos testados (p < 0, 005). Não houve diferença estatística entre PC e o GC. Conclusão: A citotoxicidade do PCS foi similar ao PH e significantemente maior do que o PS. Por outro lado, PCS e PS apresentaram um efeito genotóxico intermediário, inferior ao PH, porém superior ao PC que não teve efeito genotóxico

Hyalella azteca, an amphipod crustacean, is frequently used in freshwater toxicity tests. Since the mid-1980s, numerous organizations have collected and established cultures of H. azteca originating from localities across North America. However, H. azteca is actually a large cryptic species complex whose members satisfy both the biological and the phylogenetic species concepts. Recently, two publications reported that members of the H. azteca cryptic species complex have different toxicity responses to anions and an insecticide. In this study, four members of the H. azteca species complex were identified with DNA barcoding. The genetic variation among the four clades was consistent with interspecific distances between species. These lineages (clades 1, 3, 6, and 8) were cultured in identical conditions and monitored on a weekly basis to determine two life history traits: adult mortality and juvenile production. The large-bodied clades had significantly better survival and juvenile production compared to the small-bodied clade 3. Clade 6 had very low juvenile production and high mortality; therefore, was not included in this study. Unique culture protocols may be required for each clade to optimize growth, survival, and juvenile production in laboratory conditions. Genetic barcoding has identified only two clades in a survey of 17 laboratories. Therefore these two clades (1 and 8) were compared after exposure to copper and nickel 14-day toxicity tests. Clade 8 was 2.3-2.6 times more tolerant to copper exposure than clade 1 based on their LC50 and LC25. Similarly, clade 8 was more tolerant to nickel exposure than clade 1: LC50 was 1.8 times higher for the former. Nickel LBC50 and LBC25 were significantly different between clades by a factor of 2.1-2-8. Mortality (relative to copper concentrations in tissue), growth, and bioaccumulation responses were not significantly different based on overlapping confidence intervals. Although clades 1 and 8 are both large-bodied ecomorphs, these lineages had significantly different body mass (i.e., dry weight) after 14 days. The results of this study indicate that genetically characterized cultures of H. azteca should be used in toxicity tests.

<p>Bed bugs (<i>Cimex lectularius</i> L.) are globally important human ectoparasites. Their management necessitates the use of multiple control techniques. Plant-derived essential oils are extracts from aromatic plants that represent one of the alternative control measures for bed bug control, in addition to mechanical options and synthetic pesticides. However, there is limited information available on the efficacy and toxicology of plant essential oils against bed bugs. This project was designed with the aim to provide in-depth information on efficacy, toxicology and mode-of-action of essential oils and their insecticidal constituents in bed bugs. Initially, I evaluated topical and fumigant toxicity of fifteen essential oil components against adult male bed bugs of the Harlan strain (an insecticide susceptible strain). Neurological effects of the six most toxicologically active compounds were also determined. In both topical and fumigant bioassays, carvacrol and thymol were the most active compounds. Spontaneous electrical activity measurements of the bed bug nervous system demonstrated neuroinhibitory effects of carvacrol, thymol and eugenol, whereas linalool and bifenthrin (a pyrethroid class insecticide) produced excitatory effects. Further, I evaluated the efficacy and neurological impacts of a mixture of three neuroinhibitory compounds; carvacrol, eugenol and thymol in 1:1:1 ratio against adult male bed bugs of the Harlan strain. This mixture of monoterpenoids as well as the mixture of synthetic insecticides exhibited a synergistic affect in topical bioassays. In electrophysiology experiments, the monoterpenoid mixture led to higher neuroinhibitory effects, whereas a mixture of synthetic insecticides caused higher neuroexcitatory effects in comparison to single compounds. </p> <p>In the next objective of my dissertation, I compared the efficacy of five plant essential oils (thyme, oregano, clove, geranium and coriander), their major components (thymol, carvacrol, eugenol, geraniol and linalool) and EcoRaider^® (commercial product) between pyrethroid susceptible (Harlan) and field collected (Knoxville) bed bug populations. Initially, I found that the Knoxville strain was 72,893 and 291,626 fold resistant to topically applied deltamethrin (a pyrethroid class insecticide) compared to the susceptible Harlan strain at the LD₂₅ and LD₅₀ lethal dose levels, respectively. Synergist bioassays and detoxification enzyme assays showed that the Knoxville strain possesses significantly higher activity of cytochrome P450 and esterase enzymes. Further, Sanger sequencing revealed the presence of the L925I mutation in the voltage gated sodium channel gene. The Knoxville strain, however, did not show any resistance to plant essential oils, their major components or EcoRaider^® in topical bioassays (resistance ratios of ~ 1). In the final objective, I evaluated the efficacy of binary mixtures of above-mentioned essential oils or their major components or EcoRaider^® with deltamethrin in susceptible and resistant bed bugs. In topical application bioassays, binary mixtures of essential oils or their major components or EcoRaider^® and deltamethrin at the LD₂₅ dose caused a synergistic increase in toxicity in resistant bed bugs. Further, I studied the inhibitory effects of major essential oil components on detoxification enzyme activities (cytochrome P450s, esterases and glutathione transferases). Detoxification enzyme assays conducted using protein extracts from bed bugs pre-treated with essential oil constituents showed that these compounds significantly inhibited cytochrome P450 activity in the resistant strain, but esterase and glutathione transferase activity were unaffected. No inhibition of detoxification enzyme activities was observed in the Harlan strain bed bugs pre-treated with essential oil constituents.</p> <p>In conclusion, my dissertation research has created the foundation for utilization of natural products for bed bug management by (i) describing the efficacy of plant essential oils and their components against bed bugs, (ii) discovering synergistic interactions between essential oil components at the nervous system level, (iii) determining susceptibility of deltamethrin-resistant bed bugs to plant essential oils and their constituents and (iv) identifying synergistic effects of essential oils or their components on toxicity of pyrethroid insecticides and underlying mechanisms of this synergistic interaction. </p> <br>

El objetivo de esta tesis es estudiar el efecto del enriquecimiento por materia orgánica producida por el cultivo de peces en el mar y por los vertidos de petróleo en el medio marino. Este trabajo intenta explicar las consecuencias ecológicas de los impactos generados por este tipo de contaminación, y en diseñar de test de toxicidad para evaluar la contaminación en el sur de las costas europeas. Específicamente, en relación con el impacto de la acuicultura, los estudios de recuperación durante la Apertura de peces bentónicos de reducción de piscicultura marina, el papel de la depredación en el sistema bentónico dispersión de los residuos y los vínculos horizontales y verticales con un impacto bentónico. / The aim of this thesis is to study the effect of organic matter enrichment produced by marinefinfish farming and oil spills on the marine environment. This work is focused on elucidating theecological consequences of the impact generated by this type of pollution, and on designingtoxictity tests to evaluate pollution for southern European coasts. Specifically, related with aquaculture impact, it studies benthic recovery during open sea fish farming abatement, the role of predation in the benthic system and links horizontal and vertical waste dispersion with benthic impact.