O ácaro purpúreo, Panonychus citri (McGregor) (Acari: Tetranychidae), ainda é considerado uma praga de importância secundária nos pomares cítricos brasileiros. No entanto, nos últimos anos, sua população vem aumentando consideravelvente devido a ocorrência de surtos populacionais do ácaro que, na maioria das vezes, causam danos consideráveis nas plantas e exige que medidas de controle sejam adotadas. Neste sentido, objetivou-se com esse estudo, avaliar os níveis de suscetibilidade de diferentes espécies e variedades de plantas cítricas no desenvolvimento e reprodução de P. citri; elucidar as causas biológicas, comportamentais e demográficas responsáveis pelo aumento populacional do ácaro após a aplicação de inseticidas piretroides; demonstrar os impactos desses inseticidas sobre o predador Iphiseiodes zuluagai Denmark & Muma (Acari: Phytoseiidae) e verificar a atividade de biopesticidas à base de azadirachtin, annonin e matrine como tática adicional de controle de P. citri. Os resultados demonstraram que as espécies e variedades de plantas cítricas apresentaram diferentes níveis de suscetibilidade ao ácaro P. citri. A maior taxa de crescimento populacional do ácaro ocorreu em laranjeira Valência e limoeiro Siciliano quando comparados as laranjeiras Pera, Natal e Hamlin e tangerineira Ponkan, demonstrando que esses hospedeiros podem contribuir para o aumento populacional do ácaro. Entre os hospedeiros avaliados, laranjeira Hamlin proporcionou menor taxa de crescimento de P. citri, demonstrando que essa variedade é menos favorável ao desenvolvimento e reprodução do ácaro quando comparado aos demais hospedeiros testados. Além das diferentes taxas de crescimento do ácaro proporcionado pelas plantas hospedeiras, o uso de inseticidas piretroides e neonicotinoides também contribuem para o aumento populacional do ácaro P. citri. Com base nos resultados, o aumento na dispersão e na deterrência de alimentação e oviposição dos ácaros proporcionados pelos inseticidas piretroides, a insensibilidade dos diferentes estádios de desenvolvimento do ácaros aos inseticidas neonicotinoides e ao efeito hormese, caracterizado pelo aumento na fecundidade das fêmeas expostas principalmente aos inseticidas piretroides deltamethrin e lambda-cyhalothrin e o neonicotinoide imidacloprid foram as principais causas biológicas e comportamentais associadas ao aumento populacional de P. citri. Por outro lado, os inseticidas esfenvalerate e thiamethoxam não ocasionaram aumento significativo nos parâmetros demográficos do ácaro quando comparado aos demais tratamentos avaliados. Além das alterações na abundância populacional de P. citri, os inseticidas piretroides ocasionaram redução na sobrevivência e/ou reprodução do predador I. zuluagai, demonstrando que esses produtos podem afetar a densidade e a efetividade do predador no controle biológico de artrópodes-praga nos pomares cítricos. Diante do aumento na abundância e na frequência de surtos populacionais do ácaro P. citri após a aplicação de inseticidas piretroides e neonicotinoides e dos impactos ocasionados sobre o predador I. zuluagai novas alternativas de controle de P. citri foram investigadas no presente estudo. Os resultados demonstraram que o uso de biopesticidas à base de azadirachtin, annonin e matrine apresentaram alta atividade contra P. citri e pode ser uma importante alternativa para uso no manejo do ácaro em substituição ou rotação com acaricidas sintéticos. As informações obtidas no presente estudo contribuirão para subsidiar os programas de manejo integrado de pragas nos pomares cítricos. / The citrus red mite, Panonychus citri (McGregor) (Acari: Tetranychidae), is still considered a pest of secondary importance in Brazilian citrus groves. However, in recent years, its population has been increasing considerably due to occurrence of mite outbreaks that, in most cases, cause considerable damage to the plants and requires the adoption of control measures. Thus, the purpose of this study was to evaluate the susceptibility levels of different species and varieties of citrus plants to the development and reproduction of P. citri; to elucidate the biological, behavioral and demographic causes responsible for mite population growth after application of pyrethroids and neonicotinoids insecticides; to demonstrate the impact of these pesticides on Iphiseiodes zuluagai Denmark & Muma (Acari: Phytoseiidae) predator and to verify the activity of azadirachtin-, annonin-, and matrine-based biopesticides as additional tactic of P. citri control. The results showed that the species and varieties of citrus plants showed different susceptibility levels to P. citri mite. The highest growth rates of mite population occurred in Valencia orange and Sicilian lemon compared to Pera, Natal and Hamlin oranges and Ponkan mandarin, demonstrating that these hosts can contribute to mite population growth. Among the hosts evaluated, Hamlin sweet orange showed lower growth rate of P. citri, showing that this variety is less favorable to the development and reproduction of the mite when compared to other hosts tested. In addition to the different growth rates of the mite provided by host plants, the use of pyrethroid and neonicotinoids insecticides also contributed to the P. citri population growth. Based on the results, the increase in dispersion and feeding and oviposition deterrence of the mites provided by the pyrethroids insecticides, the insensibility of the different development stages of mites to neonicotinoids insecticides and hormesis effect, characterized by increasing the fecundity of female exposed mainly to insecticides deltamethrin and lambda-cyhalothrin pyrethroids and imidacloprid neonicotinoid were the main biological and behavioral causes associated to population growth of P. citri. On the other hand, esfenvalerate and thiamethoxam insecticides did not cause a significant increase in the demographic parameters of the mite when compared to other treatments. Besides the changes in population abundance of P. citri, the pyrethroid insecticides caused reduction in survival and/or reproduction of I. zuluagai predator, demonstrating that these products can affect density and effectiveness of the predator in the biological control of arthropod pests in citrus groves. Given the increase in abundance and frequency of outbreaks of P. citri after application of pyrethroid and neonicotinoids insecticides and of impacts caused to I. zuluagai predator new management alternatives to P. citri control were investigated in this study. The results showed that the use of azadirachtin-, annonin-, and matrine-based biopesticides had high activity against P. citri and may be an important alternative in the management of the mite in replacement or rotation with synthetic acaricides. The information obtained in this study will contribute to support the integrated pest management programs of citrus groves.

Durant leur cycle de vie, les insectes doivent faire face à différents perturbateurs pour réussir à survivre et à se reproduire. L’utilisation de plus en plus répandue des insecticides néonicotinoïdes, en raison de leur grande efficacité, a conduit à l’accumulation de résidus dans l’environnement. Ceux-ci ont certainement un effet additif toxique sur les insectes cibles. Cependant il a été montré que ces résidus peuvent aussi avoir un effet positif non désiré sur certains traits de vie des insectes ravageurs.Dans ma thèse, j’ai étudié les effets d’un insecticide néonicotinoïde sur le système olfactif d’un insecte ravageur, la noctuelle Agrotis ipsilon. Nos résultats montrent que l’exposition aigüe des mâles à des faibles doses de clothianidine modifie leurs réponses comportementales à la phéromone sexuelle: une baisse est observée à la dose 0,25 ng/insecte (<DL0) alors que la doses de 10 ng (DL20) induit une augmentation de la réponse chez les adultes naïfs ou pré-exposés à la phéromone. Cet effet biphasique à faible et très faible dose s’apparente à un effet hormétique et les modifications observées sont corrélées avec des changements de sensibilité du système olfactif central et non périphérique. Nous émettons l’hypothèse que la clothianidine agirait sur l’expression des sous-unités des récepteurs nicotiniques pour lesquels elle joue le rôle d’agoniste, changeant leur affinité pour l’acétylcholine et perturbant ou améliorant la transmission synaptique des signaux sensoriels selon la dose. Nos résultats montrent que la prise en compte d’effets de doses sublétales d’insecticides est essentielle non seulement pour les insectes non cibles, mais aussi des insectes cibles. / Insects face a multitude of environmental stresses, which they have to bypass in order to survive and reproduce. The extensive use of neonicotinoid insecticides, because of their high efficiency, leads to the accumulation of residues in the environment, which can have an additive toxic effect on target insects. However, such residues can also have unwanted positive effects on certain life traits of pest insects. In my thesis I studied the effects of a neonicotinoid insecticide on the olfactory system of the pest insect Agrotis ipsilon. Our results show that acute oral treatments of males with low doses of clothianidin modify their behavioural responses to the sex pheromone: a treatment with 0.25 ng/moth (<LD0) induces a decrease of pheromone responses whereas intoxication with 10 ng/moth (LD20) leads to an increase in the capacity of naive and pre-exposed males to locate a pheromone source as compared to controls. We propose that this biphasic effect, with low dose stimulation and very low dose inhibition is an hormeticlike effect, that is correlated with sensitivity changes within the central, but not the peripheral olfactory system. We hypothesize that the observed modifications might be due to a differential effect of clothianidin on the expression of different subunits of nicotinic acetylcholine receptors, which might change the affinity of the receptors for acetylcholine, and thus disturb or improve synaptic transmission of sensory signals as a function of the insecticide dose. Our results show that effects of sublethal doses of insecticides need to be taken into account not only for non-target, but also for target insects when evaluating pest management strategies.

Made available in DSpace on 2016-06-02T19:32:09Z (GMT). No. of bitstreams: 1 5910.pdf: 1437016 bytes, checksum: 8d574c7743c2d70359b23e9e449a0b53 (MD5) Previous issue date: 2014-06-16 / Financiadora de Estudos e Projetos / Phytoplankton cells are base of aquatic food webs, so supporting higher trophic levels and helping equilibrium maintenance in such ecosystems. As residues, nano-TiO2 particles reach aquatic environments where they pose risks to the biota. Literature is controversial about the toxicity of nanoparticles to phytoplankton in general and most data obtained so far has been based on nanoparticles concentrations well above those estimated to occur in natural aquatic environments. Because nanoparticles size vary according to its concentrations, and its toxicity and reactivity are largely dependent on its size, investigations of nanoparticles toxicity at environmentally relevant concentrations are important for the understanding of these compounds onto phytoplankton cells. On the contrary, investigations using higher concentrations can be meaningless. This study aimed at investigating the effects of titanium dioxide nanoparticles (nano-TiO2) at environmentally relevant concentrations (5.10-9 to 5.10- 6 mol L-1) on the freshwater Chlorophyceae Scenedesmus bijugus. We present data of a systematic investigation onto the effects of TiO2 nanoparticles to Scenedesmus bijugus focusing on its physiology under controlled laboratory conditions. We evaluated cell density, chlorophyll a concentrations, growth rates, photosynthetic quantum yield of PSII (continue...) / Células do fitoplâncton são a base de cadeias alimentares aquáticas, apoiando níveis tróficos superiores e ajudando na manutenção do equilíbrio desses ecossistemas. Na qualidade de resíduos, as nano-TiO2 alcançam os ambientes aquáticos onde podem impor riscos à biota. A literatura é controversa sobre a toxicidade de nanopartículas ao fitoplâncton e, em geral, a maioria dos dados têm sido obtidos com base em concentrações de nanopartículas superiores às estimadas para ambientes aquáticos naturais. Considerando que as nanopartículas variam de tamanho de acordo com sua concentração, em decorrência da formação de agregados e, que sua toxicidade e reatividade são dependentes de seu tamanho, investigações sobre a toxicidade desses materiais devem ser feitas empregando-se concentrações ambientalmente importantes, ou seja, aquelas que podem ser encontradas em ecossistemas naturais. Este estudo teve como objetivo investigar os efeitos das nanopartículas de dióxido de titânio (nano-TiO2) na clorofícea de água doce Scenedesmus bijugus em concentrações ambientalmente representativas. Nesta pesquisa, apresentamos dados de uma investigação sistemática sobre os efeitos de nano-TiO2 em amplitude de concentração de 5.10-9 a 5.10-6 mol L-1 em Scenedesmus bijugus, focando em sua fisiologia sob condições controladas de laboratório. Foram avaliadas a densidade de células, as concentrações de clorofila a, taxas de crescimento, rendimento quântico máximo do PSII (continua...)

Submitted by Allan Lopes Bacha null (allan_lb@hotmail.com) on 2017-09-13T00:04:23Z No. of bitstreams: 1 Dissertação_Allan_Lopes_Bacha.pdf: 1553490 bytes, checksum: abd4c622d45b58fff7b494c3dedd3026 (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-09-15T13:16:44Z (GMT) No. of bitstreams: 1 bacha_al_me_jabo.pdf: 1553490 bytes, checksum: abd4c622d45b58fff7b494c3dedd3026 (MD5) / Made available in DSpace on 2017-09-15T13:16:44Z (GMT). No. of bitstreams: 1 bacha_al_me_jabo.pdf: 1553490 bytes, checksum: abd4c622d45b58fff7b494c3dedd3026 (MD5) Previous issue date: 2017-07-18 / Plantas de eucalipto são sensíveis a estresses abióticos em seu desenvolvimento inicial, sendo as deficiências hídrica e nutricional dois dos mais recorrentes dentre eles. O maturador etil-trinexapac pode afetar positivamente o eucalipto, resposta caracterizada como hormese, conferindo às plantas maior tolerância aos estresses. Diante disto, objetivou-se avaliar o efeito do etil-trinexapac no crescimento inicial do eucalipto em condições de estresse hídrico e nutricional. Quatro experimentos foram conduzidos em casa de vegetação, em vasos de 15 L, sendo dois sob deficiência hídrica e dois sob deficiência nutricional (NPK). Nas duas condições, foram utilizadas duas épocas de aplicação de etil-trinexapac (antes do plantio - AP; e depois do plantio - DP) em mudas de Eucalyptus urophylla (clone I-144). Avaliou-se: trocas gasosas, teor de clorofila total, altura, diâmetro do caule, área foliar, matéria seca e relação raiz/parte aérea. Para a deficiência hídrica, conduziu-se dois experimentos simultaneamente, durante 74 dias após o plantio (DAP) do eucalipto. Os tratamentos consistiram da combinação de três doses de etil-trinexapac (0,0; 30 e 60 g i.a. ha-1) e duas condições hídricas (sem estresse e com deficiência hídrica – conduzido a 20% da capacidade de campo). Adotou-se o delineamento de blocos casualizados, em esquema fatorial 3x2, com cinco repetições. Na época de aplicação AP, a pulverização de etil-trinexapac ocorreu aos 0 DAP e na época de aplicação DP, aos 24 DAP. Houve efeito positivo para a taxa de assimilação líquida de CO2 aos 27 e 40 DAP, para o DP e AP, respectivamente. As plantas de eucalipto na ausência de deficiência hídrica apresentaram maior crescimento, independentemente da época de aplicação. Conclui-se que o etil-trinexapac não ocasionou efeito hormético no eucalipto, independentemente da condição hídrica, e as épocas de aplicação influenciaram de formas distintas as características avaliadas, não havendo efeito prejudicial em nenhuma delas. Para a deficiência nutricional, conduziu-se dois experimentos simultaneamente, durante 81 DAP. Os tratamentos consistiram da combinação de três doses de etil-trinexapac (0,0; 30 e 60 g i.a. ha-1) e quatro variações da solução nutritiva de Hoagland e Arnon (solução completa, -N, -P e -K). Utilizou-se delineamento de blocos casualizados, em esquema fatorial 3x4, com cinco repetições. Na época de aplicação AP a pulverização do maturador ocorreu aos 0 DAP, enquanto no DP, aos 33 DAP. As plantas em solução –K não diferiram das cultivadas em solução completa. O tratamento –N proporcionou maior relação raiz/parte aérea, mas resultou em menor crescimento, seguido pelo –P. Quando em –P, o etil-trinexapac teve efeito positivo para a matéria seca total (AP) e área foliar (DP). O maturador afetou positivamente algumas características fotossintéticas do eucalipto. / Eucalyptus plants are sensitive to abiotic stresses in their initial development, and water and nutritional deficiencies are two of the most recurrent among them. Trinexapac-ethyl can positively affect eucalyptus, a response known as hormesis, possible providing plants greater tolerance to stress. The objective of this study was to evaluate the effect of trinexapac-ethyl on the initial growth of eucalyptus under conditions of water and nutritional stress. Four experiments were conducted in greenhouse, in 15 L pots. Two in the water deficiency condition and two in nutritional deficiency (NPK). In both deficiency conditions, two trinexapac-ethyl application times (before planting - BP and after planting - AP) in Eucalyptus urophylla (clone I-144) were used. The following variables were evaluated: gas exchange, total chlorophyll content, height, stem diameter, leaf area, dry matter and root/shoot ratio. For water deficiency, two experiments were conducted simultaneously, during 74 days after planting (DAP) of eucalyptus. The treatments consisted of three doses of trinexapac-ethyl (0.0, 30 and 60 g i.a. ha-1) and two water conditions (no stress and water deficiency - conducted at 20% field capacity). A randomized complete block design was used in a 3x2 factorial scheme with five replications. In the BP application time, trinexapac-ethyl was sprayed at 0 DAP and in AP mode at 24 DAP. We found a positive effect for net assimilation rate at 27 and 40 DAP, for AP and BP, respectively. Eucalyptus plants in the absence of water deficiency showed higher growth, regardless application time. We concluded that trinexapac-ethyl did not cause hormesis on eucalyptus, regardless water status, and the application time influenced the characteristics evaluated in different ways, with no harmful effect in any of them. For nutritional deficiency, two experiments were conducted simultaneously, for 81 DAP. The treatments consisted of three doses of trinexapac-ethyl (0.0, 30 and 60 g i.a. ha-1) and four variations of the nutrient solution of Hoagland and Arnon: complete solution, -N, -P and -K. A randomized complete block design was used in a 3x4 factorial scheme with five replications. In the BP application time, trinexapac-ethyl was sprayed at 0 DAP, while in AP, at 33 DAP. Plants in -K solution did not differ from those grown in complete solution. The treatment -N provided a higher root/shoot ratio, but resulted in lower growth, followed by -P. In -P solution, trinexapac-ethyl had a positive effect on total dry matter (BP) and leaf area (AP). Trinexapac-ethyl positively affected some photosynthetic characteristics of eucalyptus.

O ácaro purpúreo, Panonychus citri (McGregor) (Acari: Tetranychidae), ainda é considerado uma praga de importância secundária nos pomares cítricos brasileiros. No entanto, nos últimos anos, sua população vem aumentando consideravelvente devido a ocorrência de surtos populacionais do ácaro que, na maioria das vezes, causam danos consideráveis nas plantas e exige que medidas de controle sejam adotadas. Neste sentido, objetivou-se com esse estudo, avaliar os níveis de suscetibilidade de diferentes espécies e variedades de plantas cítricas no desenvolvimento e reprodução de P. citri; elucidar as causas biológicas, comportamentais e demográficas responsáveis pelo aumento populacional do ácaro após a aplicação de inseticidas piretroides; demonstrar os impactos desses inseticidas sobre o predador Iphiseiodes zuluagai Denmark & Muma (Acari: Phytoseiidae) e verificar a atividade de biopesticidas à base de azadirachtin, annonin e matrine como tática adicional de controle de P. citri. Os resultados demonstraram que as espécies e variedades de plantas cítricas apresentaram diferentes níveis de suscetibilidade ao ácaro P. citri. A maior taxa de crescimento populacional do ácaro ocorreu em laranjeira Valência e limoeiro Siciliano quando comparados as laranjeiras Pera, Natal e Hamlin e tangerineira Ponkan, demonstrando que esses hospedeiros podem contribuir para o aumento populacional do ácaro. Entre os hospedeiros avaliados, laranjeira Hamlin proporcionou menor taxa de crescimento de P. citri, demonstrando que essa variedade é menos favorável ao desenvolvimento e reprodução do ácaro quando comparado aos demais hospedeiros testados. Além das diferentes taxas de crescimento do ácaro proporcionado pelas plantas hospedeiras, o uso de inseticidas piretroides e neonicotinoides também contribuem para o aumento populacional do ácaro P. citri. Com base nos resultados, o aumento na dispersão e na deterrência de alimentação e oviposição dos ácaros proporcionados pelos inseticidas piretroides, a insensibilidade dos diferentes estádios de desenvolvimento do ácaros aos inseticidas neonicotinoides e ao efeito hormese, caracterizado pelo aumento na fecundidade das fêmeas expostas principalmente aos inseticidas piretroides deltamethrin e lambda-cyhalothrin e o neonicotinoide imidacloprid foram as principais causas biológicas e comportamentais associadas ao aumento populacional de P. citri. Por outro lado, os inseticidas esfenvalerate e thiamethoxam não ocasionaram aumento significativo nos parâmetros demográficos do ácaro quando comparado aos demais tratamentos avaliados. Além das alterações na abundância populacional de P. citri, os inseticidas piretroides ocasionaram redução na sobrevivência e/ou reprodução do predador I. zuluagai, demonstrando que esses produtos podem afetar a densidade e a efetividade do predador no controle biológico de artrópodes-praga nos pomares cítricos. Diante do aumento na abundância e na frequência de surtos populacionais do ácaro P. citri após a aplicação de inseticidas piretroides e neonicotinoides e dos impactos ocasionados sobre o predador I. zuluagai novas alternativas de controle de P. citri foram investigadas no presente estudo. Os resultados demonstraram que o uso de biopesticidas à base de azadirachtin, annonin e matrine apresentaram alta atividade contra P. citri e pode ser uma importante alternativa para uso no manejo do ácaro em substituição ou rotação com acaricidas sintéticos. As informações obtidas no presente estudo contribuirão para subsidiar os programas de manejo integrado de pragas nos pomares cítricos. / The citrus red mite, Panonychus citri (McGregor) (Acari: Tetranychidae), is still considered a pest of secondary importance in Brazilian citrus groves. However, in recent years, its population has been increasing considerably due to occurrence of mite outbreaks that, in most cases, cause considerable damage to the plants and requires the adoption of control measures. Thus, the purpose of this study was to evaluate the susceptibility levels of different species and varieties of citrus plants to the development and reproduction of P. citri; to elucidate the biological, behavioral and demographic causes responsible for mite population growth after application of pyrethroids and neonicotinoids insecticides; to demonstrate the impact of these pesticides on Iphiseiodes zuluagai Denmark & Muma (Acari: Phytoseiidae) predator and to verify the activity of azadirachtin-, annonin-, and matrine-based biopesticides as additional tactic of P. citri control. The results showed that the species and varieties of citrus plants showed different susceptibility levels to P. citri mite. The highest growth rates of mite population occurred in Valencia orange and Sicilian lemon compared to Pera, Natal and Hamlin oranges and Ponkan mandarin, demonstrating that these hosts can contribute to mite population growth. Among the hosts evaluated, Hamlin sweet orange showed lower growth rate of P. citri, showing that this variety is less favorable to the development and reproduction of the mite when compared to other hosts tested. In addition to the different growth rates of the mite provided by host plants, the use of pyrethroid and neonicotinoids insecticides also contributed to the P. citri population growth. Based on the results, the increase in dispersion and feeding and oviposition deterrence of the mites provided by the pyrethroids insecticides, the insensibility of the different development stages of mites to neonicotinoids insecticides and hormesis effect, characterized by increasing the fecundity of female exposed mainly to insecticides deltamethrin and lambda-cyhalothrin pyrethroids and imidacloprid neonicotinoid were the main biological and behavioral causes associated to population growth of P. citri. On the other hand, esfenvalerate and thiamethoxam insecticides did not cause a significant increase in the demographic parameters of the mite when compared to other treatments. Besides the changes in population abundance of P. citri, the pyrethroid insecticides caused reduction in survival and/or reproduction of I. zuluagai predator, demonstrating that these products can affect density and effectiveness of the predator in the biological control of arthropod pests in citrus groves. Given the increase in abundance and frequency of outbreaks of P. citri after application of pyrethroid and neonicotinoids insecticides and of impacts caused to I. zuluagai predator new management alternatives to P. citri control were investigated in this study. The results showed that the use of azadirachtin-, annonin-, and matrine-based biopesticides had high activity against P. citri and may be an important alternative in the management of the mite in replacement or rotation with synthetic acaricides. The information obtained in this study will contribute to support the integrated pest management programs of citrus groves.

Made available in DSpace on 2017-07-10T14:38:31Z (GMT). No. of bitstreams: 1 Raquel_Valmorbida.pdf: 1039911 bytes, checksum: f9af83c22da423b866713b0a21b70adb (MD5) Previous issue date: 2016-03-22 / Fundação Araucária / Muvuca is a seed sowing technique of various tree species succession of distinct stages, together with species of green manure, in order to restore a degraded area. In this seeding technique, there can be biological interaction exudates, seeds and seedlings adjunct, different species of which may be mutually benefit or impaired, as germination and early development. An attempt was made in this study to investigate the allelopathic potential of exudates of Cajanus cajan seeds and Bauhinia forficata on bioindicator species (Solanum lycopersicum - tomato); and the allelopathic effect of these same exudates on forest species (Bauhinia forficata and Parapiptadenia rigida). The experiment was conducted in the laboratory, and the treatments were: control; exudates of Phase I and II of the soaking curve of C. cajan seeds and B. forficata, who underwent pH testing, electrical conductivity and phytochemical screening. The bioindicator species subjected to treatment was evaluated for germination, early growth, antioxidant enzyme activity and lipid peroxidation. Forest species subjected to the treatments were evaluated for germination and early development. According to the electrical conductivity, a greater release of exudates of precursor species in phase II of soaking. All exudates showed allelochemicals in its composition. As for the allelopathic potential, it is concluded that the exudates of C. cajan seeds and B. forficata not adversely affect germination, early growth and antioxidant enzyme activity of bioindicator, on the contrary, benefited performance these variables evaluated tomato. As for the allelopathic effect, the exudate of phase I of soaking C. cajan, and phases I and II of the seeds of B. forficata stimulated shoot length P. rigida. However, the exudate of the soaking stage II seeds of C. cajan, negatively affected the germination and initial development B. forficata. And still retarded germination of P. rigida. / Muvuca de sementes é uma técnica de semeadura de diversas espécies florestais, de distintos estágios sucessionais, juntamente com espécies de adubos verdes, com a finalidade de restaurar uma área degradada. Nesta técnica de semeadura, pode ocorrer interação biológica de exsudatos de sementes e plântulas adjuntas, das diferentes espécies, que mutuamente podem ser beneficiadas ou prejudicadas, quanto à germinação e desenvolvimento inicial. Procurou-se neste trabalho investigar o potencial alelopático de exsudatos de sementes de Cajanus cajan e Bauhinia forficata sobre espécie bioindicadora (Solanum lycopersicum - tomate); e o efeito alelopático destes mesmos exsudatos sobre espécies florestais (Bauhinia forficata e Parapiptadenia rigida). O experimento foi conduzido em laboratório, e os tratamentos foram: Testemunha; Exsudatos das Fases I e II da curva de embebição das sementes de C. cajan e B. forficata, que foram submetidos a testes de pH, condutividade elétrica e prospecção fitoquímica. A espécie bioindicadora submetida aos tratamentos foi avaliada quanto à germinação, desenvolvimento inicial, atividade enzimática antioxidante e peroxidação lipídica. As espécies florestais submetidas aos tratamentos foram avaliadas quanto à germinação e desenvolvimento inicial. Segundo a condutividade elétrica, houve maior liberação de exsudatos das espécies precursoras na fase II de embebição. Todos os exsudatos apresentaram aleloquímicos em sua composição. Quanto ao potencial alelopático, conclui-se, que os exsudatos das sementes de C. cajan e B. forficata não afetaram negativamente a germinação, desenvolvimento inicial e atividade enzimática antioxidante da planta bioindicadora, pelo contrário, beneficiou o desempenho das variáveis avaliadas do tomate. Quanto ao efeito alelopático, o exsudato da fase I de embebição de C. cajan, e das fases I e II das sementes de B. forficata estimularam o comprimento da parte aérea de P. rigida. Porém, o exsudato da fase II de embebição das sementes de C. cajan, influenciou negativamente a germinação e desenvolvimento inicial de B. forficata. E ainda retardou a germinação de P. rigida.

En Europe, le nombre de cancers est en constante augmentation et explique l’augmentation des traitements. Les bases de ces traitements sont la chimiothérapie et la radiothérapie, seules ou en association. Les chimiothérapies sont effectuées à l’aide de médicaments anticancéreux qui ont des propriétés toxiques pour les cellules. Après administration des traitements aux patients, les médicaments sont excrétés et se concentrent dans les effluents hospitaliers et les réseaux d’égouts. Bien que beaucoup de ces composés soient éliminés dans les stations d’épuration, certains sont difficilement biodégradables et sont directement rejetées dans le milieu naturel où ils représentent un risque toxique pour la flore, la faune et l’Homme. Bien que les concentrations soient faibles (ng/L - μg/L), très peu de données sont disponibles sur leurs impacts écotoxicologiques. Leur présence dans l’environnement est d’autant plus préoccupante que les produits de métabolisation sont souvent plus toxiques que la substance d’origine. L’objectif de la thèse a été d’évaluer le risque (éco)toxicologique induit par de faibles doses de médicaments rejetés seuls ou en mélanges dans les effluents hospitaliers, urbains et dans l’environnement. De par leur utilisation courante dans les traitements anticancéreux, trois molécules ont été sélectionnées pour notre étude : la ciprofloxacine (antibiotique), le tamoxifène (perturbateur endocrinien), et le cyclophosphamide (anticancéreux). Des gammes de concentrations représentatives des effluents hospitaliers, station d’épuration et de l’environnement ont été testées à l’aide de bioessais appliqués à des organismes aquatiques (V. fischeri, P. subcapitata, L. minor) et de biomarqueurs appliqués à une levure (S. cerevisiae) et des cellules humaines hépatiques et mammaires. La viabilité cellulaire (test MTS) et la génotoxicité (cassures à l’ADN et adduit à l’ADN) ont été comparés aux tests standardisés Microtox ®, Algaltoxkit F™, ainsi que d’inhibition de croissance de Lemna minor. Le potentiel perturbateur endocrinien a été évalué en parallèle à l’aide du test YES/YAS. Cette batterie de tests a ensuite été appliquée a des effluents bruts (hospitaliers, station d’épuration) pour en évaluer le potentiel (géno)toxique. Des échantillons à proximité de l’hôpital de Gérone (Espagne) ont été prélevés en sortie de l’hôpital, en entrée et en sortie de station d’épuration, pendant trois mois consécutifs. Plusieurs effets de toxicité ont été observés sur les modèles d’étude, comme notamment l’apparition de phénomènes d’hormèses sur la viabilité des cellules hépatiques exposée au tamoxifène et à la ciprofloxacine, seuls ou en mélange. Le même schéma est observé pour les mélanges avec le test Microtox®. D’autre part, l’exposition respective des cellules hépatiques et mammaires aux médicaments n’entraîne pas de cassures de l’ADN et entraine l’apparition d’adduits seulement avec le tamoxifène, alors qu’on note une augmentation dose-dépendant des cassures et des adduits à l’ADN après exposition aux mélanges. De même, une réponse positive est observée avec le test Algaltox F™. Concernant les effluents, les effets dépendent à la fois du type d’organisme et du temps d’exposition. Les tests Microtox®, Algaltox F™ et le post-marquage des adduits à l’ADN sont apparus être les pertinents pour l’analyse. Les interactions observées entre les composés mettent en avant la nécessité d’évaluer les effets des contaminants à petites doses en mélanges, à plusieurs temps d’exposition et avec différents tests. L’application d’une telle batterie de tests à des échantillons environnementaux permet de qualifier les effluents et de suivre l’efficacité de moyens d’épuration. A terme, son application pourrait permettre de mieux appréhender les risques (éco)toxiques associés aux rejets de médicaments dans l’environnement et pouvant être à l’origine de cancer secondaire chez l’Homme. / In Europe, cancers rate is constantly raising, which explain the increase in treatments. They are usually chemotherapy and radiotherapy, alone or combine. Chemotherapy is done with anticancer drugs with toxic characteristics on cells. After administer the treatments to the patients, some of the drugs are excreted in significant proportion and released in hospital and communal effluents. Even though a lot of the compounds are either removed by adsorption or bio-degradation in waste water treatment plant (WWTP), some of their are not are directly released in the environment and represent a toxic risk for aquatic organisms and the Human health. Despite low concentrations (ng/L-μg/L), few data are available about the ecotoxicological impact. The importance of chemical compounds pollutants, especially anticancer drugs, are a real concern because the metabolites of the chemicals are even more toxic than the original substance. The aim of the thesis is to develop a battery based approach to evaluate the risks induct by low doses of drugs released independently or in mixture, in hospital waste water. Because of their common use in anticancer treatment procedures, three molecules have been chosen for our study: ciprofloxacin (antibiotic), tamoxifen (endocrine disruptor), cyclophosphamide (anticancer). Concentrations were range from hospital sewers and WWTP to the environment have been tested with a battery base approach using standardized bioassays applied on aquatic organisms (V fischeri, S. subcapitata, L. minor) and biomarker applied on yeast (S. cerevisia), hepatic and mammary human cell lines. Cell viability (test MTS) and genotoxicity (DNA breaks, DNA adducts) were compared with the standardized bioassays Microtox®, Algaltoxkit F™, and Lemna minor growth inhibition. In parallel, the endocrine disruptor activity was estimated the YES/YAS assay. The battery assay was then applied to evaluate the (geno)toxicity of raw effluents (hospital, wastewater-treatment plant). Samples from the hospital of Girona (Spain) were taken got out of it from the hospital, in entrance and got out of it from water-treatment plant, during three consecutive months. Several toxic effects have been observed during this work on aquatic organisms and both human cell lines. Results show especially hormetic effect on viability of hepatic cell line exposed to ciprofloxacin and tamoxifen alone or in mixture. Same results were observed the Microtox assays after mixtures exposures. On the other hand, the individually hepatic and mammary cell exposure to the drugs doesn’t induce DNA break, and induce DNA adduct only with the tamoxifen. Furthermore, we observe a dose-dependent increasing of the DNA break and adduct if the cells are exposed in mixture. Same results were observed with Algaltox F™. Concerning the effluents, effects depending on the kind of organisms and time exposure. The Microtox ®, Algaltox F ™ and the DNA adducts post-labelling appeared to be the most relevant for the analysis. The interactions observed between drugs pinpoint the necessity to assess the effect of contaminants in low doses mixtures, at many exposure times, and using different tools. The application of this battery with environmental raw samples is in use to rank outflows toxicity and follow the WWTP efficiency, which could lead to a better understanding of the human health risks.

Stressful experiences in early life can have profound and lasting impacts on phenotypic development. In anthropogenic environments, organisms are increasingly exposed to evolutionarily novel stressors that may play a major role in shaping the phenotypic variation upon which natural selection acts. For instance, ionizing radiation persists in areas affected by nuclear reactor accidents, nuclear weapons testing, and the nuclear power production process. This thesis explored the dose-response effects of early life ionizing radiation exposure on life- history traits in the cricket (Acheta domesticus L.). Specifically, this work had two goals: (1) to examine the effects of early life radiation stress on the potential tradeoff between growth and self-maintenance, and (2) to explore the dose-dependent effects of juvenile radiation exposure on adult trait expression (particularly fecundity, offspring investment, and offspring fitness). It was found that exposure to moderate doses of radiation in early development resulted in a slower juvenile growth rate but increased survival in early adulthood, suggesting that self- maintenance was prioritized over growth. Despite the strong inhibitory effects of early life radiation on adult female body mass, age-specific fecundity was negatively impacted only at relatively high radiation doses. Crickets exposed to moderate radiation doses in early development laid larger eggs in adulthood relative to controls and these eggs had a greater hatching success, suggesting that radiation exposure had transgenerational effects on offspring performance. No noticeable effects of early life radiation exposure were detected on total and non-enzymatic antioxidant capacity or hydrogen peroxide levels in adult females. Together, this research indicates that a single, acute exposure to ionizing radiation in early life can affect phenotypic development in a complex, dose-dependent manner and that rather than being purely negative, phenotypic responses can be sustained or even enhanced. / Thesis / Master of Science (MSc) / Stressful experiences in early life can have lasting impacts on organismal development, but the extent to which early life stress is harmful is unclear. In human-dominated environments, pollutants such as ionizing radiation can be a significant form of developmental stress. This thesis explores the fitness-related effects of early life ionizing radiation exposure in the cricket, Acheta domesticus. Impacts on growth, longevity, fecundity, and offspring fitness are explored. This research indicates that early life stress can affect phenotypic development in a complex, dose-dependent manner and that rather than being purely negative, responses can be sustained or even enhanced.

La dysfonction endothéliale vasculaire constitue un marqueur précoce des maladies cardiovasculaires car l’endothélium est l’une des premières cibles des facteurs de risque cardiovasculaire. La présence d'un stress chronique engendré par les facteurs de risque cardiovasculaire sollicite les mécanismes de défense endogènes, tels que les enzymes antioxydantes, qui servent au maintien de la fonction endothéliale. L’environnement vasculaire auquel l’endothélium est exposé a un effet direct sur son fonctionnement à long terme. Certaines habitudes de vie sont ainsi associées à une bonne santé cardiovasculaire. Par exemple, la diète méditerranéenne et/ou la pratique régulière de l’exercice physique aident à maintenir une fonction endothéliale adéquate et à réduire l’incidence des maladies cardiovasculaires. D'autre part, certains gènes clés, comme le gène suppresseur de tumeurs p53, régulent plusieurs voies métaboliques importantes pour préserver l’intégrité des cellules endothéliales. Nous posons l’hypothèse que l’environnement vasculaire post-natal influence la mise en place de mécanismes de défenses endogènes tels que les enzymes antioxydantes afin de faire face à des stress plus tard dans la vie. Notre objectif global était d’évaluer les impacts d’interventions post-natales bénéfiques et d’une diminution endogène du gène suppresseur de tumeurs p53, sur la fonction endothéliale vasculaire et sur sa capacité à faire face à un stress métabolique. Dans une première étude, nous avons soumis des souris saines C57Bl/6 dès leur sevrage et jusqu’à l’âge de 9 mois, à un programme d’exercice physique volontaire (course dans une roue) ou à un antioxydant (catéchine), comparé à un groupe de souris sédentaires et sans antioxydant. Puis les interventions ont été stoppées et une diète riche en gras a été introduite, ou non, pour une période de 3 mois; les souris ont été sacrifiées à l'âge de 9 ou 12 mois. Nous avons observé que l’exercice a protégé les cellules endothéliales des effets délétères induits par la diète riche en gras en préservant la fonction endothéliale par le maintien d’un profil rédox sain et en évitant la hausse de l’inflammation. La catéchine a maintenu la fonction endothéliale aortique, mais n’a pas prévenu le profil inflammatoire en présence de la diète riche en gras. Finalement, chez les souris sédentaires, la fonction endothéliale a été détériorée en présence de la diète riche en gras, sans indice d’inflammation vasculaire. Dans une seconde étude, des souris partiellement déficientes en p53 (p53+/-) et contrôles C57Bl/6 ont été exposées à la même diète riche en gras à partir de 3 mois et ce jusqu’à l’âge de 6 mois. Notre raisonnement était basé sur la démonstration que p53 est un régulateur de l’expression des enzymes antioxydantes in vitro. Chez les souris p53+/-, les cellules endothéliales ont été protégées du stress induit par l’hypercholestérolémie engendrée par la diète riche en gras. Cependant, chez les souris p53+/- cette protection pourrait être secondaire à un métabolisme accru des acides biliaires, qui en prévenant la hausse de cholestérol, protègerait indirectement l'endothélium. Nous avons donc pu démontrer l’importance de l’environnement vasculaire sur la fonction endothéliale. La diète riche en gras a stimulé certains mécanismes de défense vasculaires tels que la voie des EDHF et la superoxyde dismutase afin de maintenir la fonction endothéliale malgré les conditions pro-athérosclérotiques. Nous avons observé que l’exercice et la catéchine influencent différemment l’endothélium malgré leurs capacités antioxydantes. Ces études soulignent la sensibilité de l’endothélium aux changements dans l’environnement vasculaire. En accord avec le vieillissement de la population et la progression des maladies cardiovasculaires, la proportion de personnes ayant une dysfonction endothéliale augmente. Ainsi, une meilleure compréhension des mécanismes ou d’interventions qui permettent le maintien de la fonction endothéliale à long terme s’avère utile. / Endothelial dysfunction is an early marker of atherosclerosis and cardiovascular diseases. Cardiovascular risk factors generate a chronic stress, challenging endogenous defense mechanisms that are critical to maintain endothelial function, such as antioxidant enzymes. The vascular environment impacts the integrity and long-term function of endothelial cells. Thus, a healthy lifestyle is beneficial for cardiovascular health: regular physical training and/or a Mediterranean diet are associated with the maintenance of endothelial function and a lower incidence of cardiovascular diseases. On the other hand, some key genes such as tumor suppressor gene p53, are known to regulate numerous cellular functions that are necessary to maintain endothelial cells integrity. We hypothesized that the post-natal vascular environment impacts the development of endogenous cellular defense mechanisms such as antioxidant enzymes, in order to protect against vascular stress that will occur later in life. Our major goal was to evaluate the impact of post-natal interventions and endogenous reduction of p53 expression on vascular endothelial function and its capacity to resist against a metabolic stress. In our first study, healthy C57Bl/6 mice were exposed from weaning to the age of 9 months to physical voluntary training (running wheel) or to the antioxidant catechin, and were compared to physically inactive mice that did not receive catechin. Then, exercise and catechin were stopped and mice were subjected to a regular or a high fat diet for 3 months; mice were sacrificed either at the age of 9 or 12 months. In trained mice, we observed that exercise prevented endothelial dysfunction and inflammation induced by the high fat diet. In catechin-treated mice, aortic endothelial function was maintained despite exposure to the high fat diet, but an inflammatory profile was observed. In physically inactive mice, the high fat diet induced endothelial dysfunction without inflammation. In our second study, mice partially deficient in p53 (p53+/-) were exposed to the same high fat diet from 3 to 6 months of age. Our rationale was based on the discovery that in vitro, p53 regulates antioxidant enzymes gene expression. In p53+/- mice, endothelial cells were protected from the stress of hypercholesterolemia induced by the high fat diet. However, this endothelial protection could be linked with an unexpected enhanced bile acid metabolism in p53+/- mice: low endogenous expression of p53 prevents the rise in plasma total cholesterol when fed a high fat diet, indirectly protecting the endothelium. In summary, we were able to demonstrate the importance of the vascular environment on endothelial function. The pro-atherosclerotic environment induced by the high fat diet stimulated vascular defense mechanisms, as observed by the activation of the compensatory EDHF pathway and superoxide dismutase activity, to maintain an adequate endothelial function. We observed that exercise and catechin had a different impact on the endothelium despite their antioxidant properties. These studies demonstrate the sensitivity of the endothelium to changes within the vascular environment. As the population is aging and the incidence of cardiovascular diseases is increasing, endothelial dysfunction will be more frequent. Therefore, a better comprehension of mechanisms or interventions that can protect endothelial function can only be beneficial.

Diese Dissertation ist ein Beitrag zum Forschungsfeld der Stressökologie, im Spe-ziellen der Verhaltensökotoxikologie. Das spontane lokomotorische Verhalten der Fischarten Danio rerio und Leucaspius delineatus wurde unter sublethaler Expo-sition mit dem Cyanobakterientoxin Microcystin-LR (MC-LR) und dem Xenobio-tikums 2.4.4`-Trichlorobiphenyl (PCB 28) quantifiziert. Die Schwimmgeschwin-digkeit und Anzahl der Wendungen wurden kontinuierlich mit einem automati-schen Video-Monitoringsystem unter Laborbedingungen aufgezeichnet. In Hin-blick auf zyklische Aspekte wurden die Verhaltensanalysen mit chronobiologi-schen Methoden kombiniert. Hiermit wurde gezeigt, dass MC-LR und PCB 28 zu signifikanten Effekten in Verhalten und Aktivitätsrhythmik beider Fischarten führten. Höhere Konzentrati-onen beider Untersuchungssubstanzen verursachten eine deutliche Aktivitätsredu-zierung bei Danio rerio und Leucaspius delineatus. Einige der festgestellten Do-sis-Wirkungsbeziehungen entsprechen der Hormesistheorie, z. B. war bei geringe-ren MC-LR Konzentrationen ein Aktivitätsanstieg und bei höheren ein Aktivitäts-abfall beider Fischarten zu verzeichnen. Die Exposition mit MC-LR und PCB 28 verringerte bei beiden Testfischarten die Synchronisation der Aktivität mit dem Zeitgeber Licht. Dies führte bei beiden Fischarten zu einer Phasenverschiebung. Bei Leucaspius delineatus war unter dem Einfluss von MC-LR eine Phasenumkehr zu verzeichnen, die Fische wechselten von Tag- zu Nachtaktivität. Die Cosinor Analyse zeigte Dosis abhängige Veränderungen der circadianen Rhythmen der Schwimmaktivität (z.B. MESOR, Akrophase) unter Einfluss von MC-LR und PCB 28 an. Die Power Spektral Analyse indizierte für beide Fischar-ten unter Einwirkung von MC-LR and PCB 28 eine reduzierte Dominanz des cir-cadianen Rhythmuspeaks. Da die registrierten Unterschiede in der Reaktion beider Fischarten auf MC-LR und PCB 28 eher gering waren, sind Ergebnisse der Art Danio rerio, die häufig in Toxizitätstests verwendet wird, mit denen der einheimischen Art Leucaspius deli-neatus vergleichbar. Die Ergebnisse belegen, dass Verhaltensuntersuchungen in Kombination mit chronobiologischen Auswertemethoden eine sensitive und zuverlässige Abschät-zung des Gefährdungspotentials von Substanzen sowohl auf dem Gebiet der Öko-toxikologie als auch für Biomonitoring ermöglichen. / This thesis contributes to the field of stress ecology specifically behavioural ecotoxicology. The spontaneous locomotor behavior of two fish species Danio re-rio and Leucaspius delineatus was recorded and quantified continuously under sublethal exposure to the cyanotoxin microcystin-LR (MC-LR) and the xenobioti-cum 2.4.4`-trichlorobiphenyl (PCB 28). By using an automated video-monitoring system, the swimming velocity and the number of turns were assessed under labo-ratory conditions. For analysing cyclic aspects basic behavioural analyses were combined with chronobiological procedures as cosinor analysis and power spec-tral analysis. Both MC-LR and PCB 28 acted as stressors and caused significant changes in the behaviour and circadian activity rhythms of Danio rerio as well as Leucaspius de-lineatus. So for both species elevated concentrations of the stressors led to a re-duction of their activity. Some dose-responses correspond to the hormesis theory, e.g., there was an increase of daytime activity at lower MC-LR concentrations and a decrease at elevated concentrations of MC-LR for both species. A degree of desynchronisation of activity to the zeitgeber light, which led to a phase shift was caused by the chemicals in both fish species. In Leucaspius de-lineatus this shift was so drastic that this species reversed their significant diurnal activity and became nocturnal under the influence of MC-LR. The Cosinor analysis revealed MC-LR- and PCB 28-induced, dose-dependent al-terations of the circadian rhythms of activity (e.g., MESOR, acrophase). The power spectral analysis showed that the dominance of the circadian rhythmic peak (of 24 h) was reduced under MC-LR and PCB 28 for both species. Since the observed differences in the reactions of both species to MC-LR and PCB 28 were rather small, the results of the species Danio rerio which is widely used for environmental risk assessment tests, are comparable to those of the native European species Leucaspius delineatus. The findings of this study proved that the basic behavioural analyses combined with chronobiological procedures could be valuable tools for the study of stressful or even harmful environmental factors in the field of ecotoxicology as well as for biomonitoring.