Influence of Chironomus riparius (Diptera, Chironomidae) and Tubifex tubifex (Annelida, Oligochaeta) on oxygen uptake by sediments. Consequences of uranium contamination

The diffusive oxygen uptake (DOU) of sediments inhabited by Chironomus riparius and Tubifex tubifex was investigated using a planar oxygen optode device, and complemented by measurements of bioturbation activity. Additional experiments were performed within contaminated sediments to assess the impact of uranium on these processes. After 72 h, the two invertebrate species significantly increased the DOU of sediments (13–14%), and no temporal variation occurred afterwards. Within contaminated sediments, it was already 24% higher before the introduction of the organisms, suggesting that uranium modified the sediment biogeochemistry. Although the two species firstly reacted by avoidance of contaminated sediment, they finally colonized it. Their bioturbation activity was reduced but, for T. tubifex, it remained sufficient to induce a release of uranium to the water column and an increase of the DOU (53%). These results highlight the necessity of further investigations to take into account the interactions between bioturbation, microbial metabolism and pollutants

Bioavailability of pesticides in freshwater sediments

In ecological risk assessment standardized sediment toxicity tests are used to predict the hazard of chemicals for sediment-living organisms. Feeding is a prerequisite in these long-term tests to avoid starvation of test organisms. Therefore, added food particles may act as vectors for the test compound. The importance of food particles as vectors, however, is dependent on several factors, for example, sorption and major uptake routes. In this thesis, laboratory experiments on the importance of pesticide sorption and uptake routes for the bioavailability to the midge Chironomus riparius in sediment toxicity test setups were performed. A feeding selectivity study showed that larvae almost exclusively fed on added food particles, and highly neglected sediment particles. Additionally, experiments on the sorption of the insecticide lindane, showed that food and peat particles (used in artificial sediment) efficiently sorbed lindane (>95% after 48 h). The binding strength of lindane was weak, facilitating particulate uptake. However, the uptake from dissolved lindane was higher than the uptake from particles. From this we concluded that toxicity may be underestimated in spiked-sediment scenarios, where hydrophobic pesticides sorb to the sediment and larvae to a large extent feed on uncontaminated food particles. Conversely, in a spiked-water scenario, the food particles may act as vector, resulting in a facilitated particulate uptake, in addition to the uptake from water. Sediment organic matter affects sorption, and thus bioavailability of pesticides. Pyrethroid toxicity was much higher in artificial sediment than in a natural sediment, indicating the simplicity and shortcomings of using artificial sediments. Interestingly, the sediment quality highly affected bioavailability in spiked-water. For example, C. riparus larvae in sediments with low organic matter content and exposed to spiked-water pyrethroids, showed lower survival, slower development, and increased adult size, than those in sediments with higher organic matter. The pyrethroid deltamethrin, showed an LC50-value (28 d) for C. riparius larvae in artificial sediment of 16 pg/L and 11 µg/kg for water- and sediment exposures, respectively, i.e. toxic effects occurred at concentrations lower than the detection limits for high-tech analytical methods. This thesis contributes to a wider understanding of processes affecting bioavailability in freshwater sediments, and in particular in standardized sediments used in toxicity testing. The understanding of test compound sorption and bioavailability is crucial for sound interpretations of toxicity tests and for the general credibility of such tests

Toxicidade de pesticidas para Chironomus riparius: alterações no proteoma, marcadores bioquímicos e respostas individuais

The application of pesticides in agricultural fields leads to inevitable contamination of adjacent freshwater systems, representing a serious threat to non-target aquatic invertebrate communities. The study of the impact of these stressors on ecologically relevant species is crucial for risk assessment. Traditionally, toxicity testing focuses on organism and population-level responses (e.g. mortality, growth, behavior, and reproduction). However, these responses are often preceded by changes at lower levels of biological organization. In this sense, there is a need to develop sensitive tools that can be used to predict ecological adverse effects of sub-lethal concentrations of pesticides. Assessing sub-organismal endpoints may therefore provide early indicators of pesticide exposure and their possible impacts on natural populations. In this thesis, larvae of Chironomus riparius (Meigen) were exposed to four insecticides with distinct modes of action: amitraz, spinosad, indoxacarb, and fipronil, and their effects evaluated in terms of life-history responses using standard laboratory ecotoxicological tests, and at biochemical level by monitoring specific oxidative stress, neuronal, and energy metabolism biomarkers. Moreover, the effects of spinosad, indoxacarb, and fipronil were assessed at the molecular level using proteomic tools, to determine if proteomics and biochemical biomarkers can be used as reliable and sensitive tools in ecological risk assessment. The results presented here indicate that environmentally relevant concentrations of the insecticides tested can significantly affect several C. riparius life-history traits, with reductions in the larval growth and impairment of emergence endpoints observed for all compounds tested, which ought to compromise the ecological integrity of freshwater ecosystems. At the biochemical level, very distinct responses were observed for each pesticide, probably due to their distinct modes of action. Nonetheless, evidences of high metabolic costs (as indicated by the increase of electron transport system (ETS) and/or lactate dehydrogenase (LDH) activities) were observed for all insecticides, which are probably associated with the activation of antioxidant defenses and detoxification processes. Additionally, evidences of oxidative damage were found in C. riparius larvae under exposure to amitraz and spinosad, as indicated by the increase in lipid peroxidation (LPO) levels. At the proteome level, no significant changes were found in C. riparius proteome between exposed and non-exposed larvae for the concentrations of indoxacarb tested. Fipronil exposure induced alterations in the expression of globins, cytoskeleton and motor proteins, as well as in proteins involved in protein synthesis. Exposure to spinosad resulted in alterations in globins, actin, and cuticle proteins’ expression. These changes observed at the proteome level revealed potential mechanisms of action that lead to the effects observed at the individual level. The potential of C. riparius globins expression in environmental monitoring studies has been previously stated and are here sustained. The generalized downregulation of these proteins observed under exposure to spinosad and fipronil may be related to the toxic effects of these insecticides. This study highlights the importance of complementing standard ecotoxicological approaches with biochemical and molecular tools in an integrative manner. The analyses of biochemical biomarkers and of the proteome can be useful in risk assessment, contributing to the knowledge of the sub-lethal effects of pesticides, thus aiding the comprehensive and mechanistically understanding of the mechanisms that lead to higher level responses. It is also demonstrated that the pesticides tested here pose a potential risk to non-target aquatic invertebrates, and therefore their application near freshwater systems should be reviewed. Chironomus riparius, a model organism in aquatic toxicology, is also presented as a promising model organism for environmental proteomics.O uso de pesticidas em campos agrícolas resulta na inevitável contaminação dos sistemas de água doce adjacentes, representando uma séria ameaça para as comunidades de invertebrados aquáticos não alvo. O estudo do impacto destes compostos em espécies ecologicamente relevantes é crucial para a avaliação de risco. Tradicionalmente, os testes ecotoxicológicos baseiam-se em respostas ao nível do organismo e da população (ex. mortalidade, crescimento, comportamento e reprodução). No entanto, estas respostas observadas ao nível do organismo e população são usualmente precedidas por alterações nos níveis mais baixos de organização biológica. Nesse sentido, existe a necessidade de desenvolver ferramentas sensíveis que possam ser usadas para prever potenciais efeitos adversos ecológicos de concentrações sub-letais de inseticidas. A avaliação de efeitos ao nível subindividual pode assim fornecer informação prévia da exposição a pesticidas e os seus possíveis impactos em populações naturais. Nesta tese, larvas da espécie modelo em ecotoxicologia Chironomus riparius (Meigen) foram expostas a quatro inseticidas com diferentes modos de ação: amitraz, spinosad, indoxacarb e fipronil, e os seus efeitos avaliados em termos de respostas do ciclo de vida utilizando testes ecotoxicológicos padronizados, e ao nível bioquímico monitorizando biomarcadores específicos de stress oxidativo, neurotoxicidade e metabolismo energético. Além disso, os efeitos do spinosad, indoxacarb e fipronil ao nível molecular foram avaliados usando ferramentas de proteómica, com o objetivo de determinar se a proteómica e os marcadores bioquímicos podem ser ferramentas sensíveis na avaliação de risco ecológico. Os resultados aqui apresentados indicam que concentrações ambientalmente relevantes dos pesticidas testados, podem comprometer significativamente vários indicadores do ciclo de vida de C. riparius. Foram observadas reduções no crescimento larval e alterações nos parâmetros relacionados com a emergência dos insectos em resposta à exposição a todos os inseticidas testados, o que pode comprometer a integridade ecológica dos ecossistemas de água doce. Ao nível bioquímico, foram observadas respostas muito distintas para cada pesticida, provavelmente devido aos seus diferentes modos de ação. No entanto, foram observados indícios de elevados custos metabólicos (indicados pelo aumento das atividades da cadeia transportadora de eletrões (ETS) e/ou da enzima lactato desidrogenase (LDH) para todos os inseticidas. Estes aumentos estão provavelmente relacionados com a ativação de mecanismos de defesa antioxidantes e de processos de destoxificação. Além disso, foram observados indícios de dano oxidativo em larvas expostas a amitraz e spinosad, indicado pelo aumento nos níveis de peroxidação lipídica (LPO). Ao nível do proteoma, não foram observadas alterações significativas nas larvas expostas a indoxacarb em comparação com larvas não expostas. A exposição ao fipronil causou alterações na expressão de globinas, de proteínas motoras e do citoesqueleto, bem como em proteínas envolvidas na síntese proteica. A exposição ao spinosad resultou em alterações na expressão de globinas, actinas e de proteínas da cutícula. Estas alterações observadas ao nível do proteoma revelaram potenciais mecanismos de ação que levam aos efeitos observados ao nível do organismo. O potencial da expressão das globinas de C. riparius em estudos de monitorização ambiental foi previamente afirmado e é aqui sustentado. O decréscimo generalizado observado na expressão destas proteínas sob exposição ao spinosad e ao fipronil pode estar relacionado com os efeitos tóxicos destes inseticidas. Esta tese destaca a importância de complementar de uma forma integrada os ensaios ecotoxicológicos padronizados com ferramentas bioquímicas e moleculares. A análise de marcadores bioquímicos e do proteoma pode ser útil na avaliação de risco, contribuindo para o conhecimento dos efeitos sub-letais dos pesticidas, auxiliando na compreensão dos mecanismos que conduzem às respostas observadas nos níveis mais elevados de organização biológica. Este estudo revela também que os pesticidas testados representam um risco para os invertebrados aquáticos não alvo, e, portanto, a sua aplicação próxima de sistemas de água doce deve ser revista. Chironomus riparius, um organismo modelo em toxicologia aquática, é também aqui apresentando como um modelo promissor em estudos de proteómica ambiental.Programa Doutoral em Biologi

Three-Dimensional Time-Resolved Trajectories from Laboratory Insect Swarms

Aggregations of animals display complex and dynamic behaviour, both at the individual level and on the level of the group as a whole. Often, this behaviour is collective, so that the group exhibits properties that are distinct from those of the individuals. In insect swarms, the motion of individuals is typically convoluted, and swarms display neither net polarization nor correlation. The swarms themselves, however, remain nearly stationary and maintain their cohesion even in noisy natural environments. This behaviour stands in contrast with other forms of collective animal behaviour, such as flocking, schooling, or herding, where the motion of individuals is more coordinated, and thus swarms provide a powerful way to study the underpinnings of collective behaviour as distinct from global order. Here, we provide a data set of three-dimensional, time-resolved trajectories, including positions, velocities, and accelerations, of individual insects in laboratory insect swarms. The data can be used to study the collective as a whole as well as the dynamics and behaviour of individuals within the swarm

Mite Diet Sequences Obtained by High Throughput Sequencing of Gut Contents of Freshly Collected Water Mites

Cytochrome oxidase subunit I (COI) barcode sequences in this file were obtained from gut DNA extracted from 54 freshly collected water mites, comprising 21 Lebertia quinquemaculosa , 30 Lebertia davidcooki , 1 Limnesia , and 2 Arrenurus specimens. Methods and other details about these sequences are described in a paper by the same authors in a submitted publication (2021: URL to be given here when published). Data on collection locations, primers (mLep and LCOI), amino acid translations, etc. are included in corresponding sequences uploaded to GenBank. The right column below contains additional notes on naming the taxa of the sequences that were not included in the GenBank annotation. These notes include the highest percentage identity to a previous sequence in GenBank as determined by BLASTN in June 2018. The FASTA file name given here includes the Accession ID, followed by the best match taxon (at an appropriate taxonomic level, dependent on the percent identity, as described in the notes in the right-hand column), the phrase water mite diet isolate , a specific RamLab sequence identifier of the sequence, and then the COI gene description. Accession IDs of sequences uploaded to GenBank begin with MW; other sequences begin with RL and a RamLab sequence identifier. The RamLab sequence identifier in the FASTA name includes information as follows: RamLab ID number-location and date of collection with three location letters (e.g., BHL stands for Blue Heron Lagoon) and the date usually in a 6-character format of MMDDYY-information on the location of sequence on the Illunina sequencing plate-and a 4- to 6-character identifier of the mite species (Lq=L. quinquemaculosa ; Ldc=L. davidcooki ; Lim=Limnesia ; Arr=Arrenurus ) and the animal number in that series of experiments (2 digits)

Avaliação dos efeitos ecológicos de filtros ultravioleta em ecosistemas lóticos portugueses

Organic UV-filters are chemicals present in several personal care products and on other materials (plastics, paints, etc) to protect against ultraviolet radiation. The use of these compounds has been increasing throughout the years and, consequently, they are now considered global emergent contaminants of the aquatic environment. In fact, UV-filters have been frequently found in river and lake waters but to date, only scarce information exists about their effects and it is mostly based in acute or chronic toxicity data for a limited number of species. Due to their physico-chemical properties, UV- filters accumulate in biota and sediments and are expected to co-occur together with other persistent contaminants. Moreover, and despite uncertainties about their specific mode of action, research has shown endocrine disruption caused by exposure to organic UV-filters in several species. Investigation on the ecological effects of organic UV-filters is urgent for a correct environmental risk assessment. For that, studies should be focused on ecotoxicological data from benthic organisms considering relevant endpoints and exposure scenarios including binary mixtures. This will allow to address potential long-term as well as community and ecosystem level effects of organic UV-filters. Thus, the purpose of this thesis was to evaluate the effects of UV-filters using an integrated approach combining sub-organismal, organismal, population and community level responses, aiming to produce comprehensive and sound ecotoxicological data for freshwaters. With that purpose four specific objectives were proposed: i) assess the effects of selected UV-filters (Benzophenone-3 (BP3); 3-(4-methylbenzylidene)camphor (4-MBC) and Octocrylene (OC)) on two aquatic species, the dipteran Chironomus riparius (a model organism widely used in ecotoxicological assays) and the trichopteran Sericostoma vittatum (an endemic species of Iberian peninsula), both aquatic detritivores with an important role in the streams and rivers; ii) assess the effects of binary mixtures of different UV-filters and UV- filters combined with an insect repellent, DEET (N,N-diethyl-3-methylbenzamide) in C. riparius reproductive traits ; iii) assess the long-term effects of BP3 exposure over two consecutive C. riparius generations and iv) assess the effects of 4-MBC in the structure of macroinvertebrates community and functioning of ecosystem using a mesocosms approach. Obtained results showed that selected UV-filters indeed caused deleterious effects on both insect species at environmental relevant concentrations with reductions in the growth, feeding and development rates. At the sub-organismal level, C. riparius and S. vittatum showed different patterns of response to UV-filters exposure and, despite evidences of metabolic costs related with detoxification, no evidences of oxidative stress or neurotoxicity were found. Additionally, results obtained in mixture exposures showed synergistic interactions for some C. riparius reproduction related traits for binary mixture of BP3 and 4-MBC as well as for mixture containing BP3 and DEET. These suggest that individual chemical testing can underestimate toxicity of organic UV-filters under natural conditions. Concerning C. riparius long-term multigenerational exposure to BP3, our results showed that C. riparius fertility is strongly reduced by BP3 exposure being the filial (F1) generation more sensitive than the parental (P) generation. Moreover, the F1 generation exposed under control conditions but whose parents were exposed to BP3 showed to be affected. All these results indicate carry-over effects, pointing out to possible epigenetic/transgenerational effects. Finally, community ecotoxicological experiments using artificial streams (mesocosms) showed no effects of 4-MBC on the structure of macroinvertebrate community nor on leaf litter decomposition. However, primary production was strongly reduced due to 4-MBC exposure. These results, although not in agreement with the previous laboratory assays performed with detritivore species, suggest potential bottom up indirect effects caused by the presence of organic UV-filters in sediments. In conclusion, the results obtained in this work suggest that UV-filters present risk to freshwater benthic invertebrate communities and reinforce the need of using complex and higher tier ecotoxicity studies to a better environmental risk assessment of these emergent contaminants.Os Filtros ultravioleta (filtros UV) são compostos químicos usados em vários produtos de higiene pessoal e materiais (plásticos, tintas, etc.) de modo a proteger contra danos causados pela radiação ultravioleta. O uso destes compostos tem vindo a aumentar nos últimos anos sendo já considerados contaminantes emergentes dos ecossistemas aquáticos. De facto, estes compostos têm vindo a ser detetados em lagos e rios, mas até à data pouca informação existe sobre os seus efeitos nos ecossistemas aquáticos, sendo que os estudos existentes se centram em ensaios laboratoriais e num reduzido número de espécies. Contudo, devido às suas propriedades físico-químicas é esperado que os filtros UV acumulem nos sedimentos e nos organismos e coocorram com outros contaminantes. No entanto, embora o modo de ação dos filtros UV não seja totalmente conhecido, estudos prévios têm mostrado disrupção endócrina causada pela exposição a estes contaminantes em organismos aquáticos. Posto isto, uma avaliação dos efeitos ecotoxicológicos dos filtros UV é urgente para uma correta avaliação do risco ambiental. Assim, os estudos ecotoxicológicos devem-se focar em organismos bentónicos considerando parâmetros e cenários de exposição relevantes, o que nos permitirá determinar potenciais efeitos a longo prazo assim como efeitos ao nível das comunidades dos ecossistemas. Este trabalho propôs avaliar e perceber quais os efeitos dos filtros UV em organismos e comunidades de água doce usando uma abordagem baseada em diferentes níveis de organização biológica e tendo como intuito gerar dados ecotoxicológicos robustos e mais abrangentes. Para tal, foram estabelecidos vários objetivos específicos: 1) avaliar os efeitos de diferentes filtros UV (Benzofenona-3 (BP3); 3-(4-methylbenzylidene)camphor (4-MBC) e Octocrileno (OC)) usando duas espécies de invertebrados aquáticos detritívoros, o díptero Chironomus riparius (uma espécie modelo amplamente utilizada em ensaios ecotoxicológicos) e o tricóptero Sericostoma vittatum (uma espécie endémica da Península Ibérica), ambos com grande importância nos ecossistemas lóticos portugueses; 2) avaliar os efeitos de misturas de diferentes filtros UV, e de filtros UV juntamente com um repelente de insetos (N,N-diethyl-3-methylbenzamide -DEET) em parâmetros relacionados com a reprodução de C. riparius; 3) avaliar os efeitos multigeracionais decorrentes de exposição a BP3 em C. riparius; 4) usando sistemas de rios artificiais avaliar os efeitos do 4-MBC na estrutura e funcionamento dos ecossistemas. Os resultados obtidos mostraram que, de facto, os filtros UV selecionados causaram efeitos tóxicos em ambas as espécies usadas a concentrações ambientalmente relevantes, observando-se reduções ao nível do crescimento e das taxas alimentares assim como atrasos no desenvolvimento. Ao nível subcelular C. riparius e S. vittatum mostraram diferentes padrões de resposta à exposição aos filtros UV e apesar de se observarem custos metabólicos relacionados com os processos de destoxificação, não se observaram efeitos em termos de stress oxidativo nem neurotoxicidade. Adicionalmente, os resultados obtidos na exposição às misturas entre químicos revelaram interações sinergísticas para alguns parâmetros reprodutivos de C. riparius para as ambas as misturas testadas (BP3 – 4-MBC; BP3 - DEET). Estes resultados sugerem que exposições individuais aos filtros UV podem subestimar a toxicidade destes contaminantes em condições naturais. Nos ensaios multigeracionais observou-se também uma forte redução na fertilidade de C. riparius expostos a BP3. Observou-se ainda que a segunda geração foi mais afetada que a geração parental mesmo se mantida em condições controlo, mas cujos pais tinham sido previamente expostos a BP3. Todos estes resultados indicam que a BP3 provoca efeitos latentes na geração parental que são posteriormente observados nos descendentes, apontando assim para efeitos epigenéticos/transgeracionais. Por último, nos ensaios dos mesocosmos, não foram observados efeitos do 4-MBC na estrutura das comunidades de macroinvertebrados nem na degradação da matéria orgânica, em contraste com o forte efeito observado na produção primária. Estes resultados, embora não concordando com os resultados obtidos em ensaios laboratoriais sugerem potenciais efeitos indiretos da presença dos filtros UV nos sedimentos. Em conclusão, os resultados obtidos nesta tese indicam que os filtros UV apresentam risco para os organismos bentónicos e reforçam a necessidade de utilizar abordagens integradas com maior relevância ecológica para uma melhor avaliação do risco ambiental destes contaminantes emergentes.Programa Doutoral em Biologi

The nucleolar organizer of the salivary gland polytene chromosomes as a measure of recent growth in laboratory-reared and field-collected Chironomus Spp. (Diptera: Chironomidae) larvae

Midge larvae possess giant polytene chromosomes. Genes on these chromosomes undergoing transcription are visible as puffs. The nucleolar organizer (NOR), an especially large puff, shrinks when a larva is stressed. Two feeding experiments were conducted to examine how NOR size changes as a function of Chironomus riparius growth. NOR size was linearly related to an individual\u27s recent growth rate, independent of its body size. Chironomids were collected from wetlands constructed with oil sands mine water and tailings (OSPM), and reference wetlands to evaluate the utility of the NOR as a field-based measure of larval growth and condition. Small larvae

Effects of propagule pressure, environmental factors, and climate change on success and impacts of benthic aquatic invasions

Nonnative species introductions are linked to anthropogenic drivers, including transport of species into novel habitat(s), changes in local environmental factors that may facilitate invasions, or large-scale shifts in abiotic conditions with climate change. In freshwater ecosystems, transport of nonnative species often occurs via boats (e.g., trailers, boat props, bilge water). Environmental factors include differences in watershed land use and water quality (e.g., transparency, nutrient and chlorophyll-a concentrations, pH, and conductivity). Climate change may increase water temperatures, affecting lake stratification, and causing low or pulsed dissolved oxygen concentrations. Both understanding factors causing spatial variability of invasive species and assessing potential negative impacts to ecosystems are important. For example, some invasive species are more tolerant or adapted to increased water temperature or lower dissolved oxygen, and may persist in such environments, while natives decline. This research assessed propagule pressure, environmental factors, and climate change, focusing on benthic macrophytes and macroinvertebrates. First, propagule risk (proxy for propagule pressure, capturing extent to which invasives might be introduced from different populations) and environmental conditions were correlated with richness and abundance of invasive macrophytes in 20 lakes; results suggest that propagule risk was the driver of invasive macrophytes in the lake communities studied. Negative impacts on macrophyte communities from the introduction of a dominant macroalgae, Nitellopsis obtusa, were also evaluated. Increased abundance of N. obtusa was correlated with reductions in total and native macrophyte richness, a pattern consistent across depths, suggesting that N. obtusa can displace species in lake communities. Finally, bioenergetics and nutrient/trace metal sediment release from the nonnative oligochaete, Branchiura sowerbyi, were compared to those of native benthic invertebrates (Hexagenia and Chironomus riparius) under various water temperature and DO concentrations, to understand ecosystem implications of B. sowerbyi’s introduction. Results suggest that B. sowerbyi may be more tolerant of conditions under predicted future climate, and thus may spread and become more dominant in benthic communities, with implications for sediment nutrient and contaminant dynamics. Results of this research offer insight into ecological processes involved in benthic aquatic invasions, including factors that may lead to their success, and the consequences for ecosystems and native species once established

O papel de presas contaminadas no comportamento e fisiologia das planárias na investigação de microplásticos

Microplastics (1 μm to 1 mm) have become a concerning topic in the scientific community when associated with poor plastic waste management. Although the presence and abundance of these particles in freshwater environments have been previously reported, there is still an insufficient number of studies providing information about the potential trophic transfer and its consequences on freshwater benthic organisms. This work aimed to evaluate the behavioural and physiological responses of planarian Girardia tigrina after consuming contaminated prey - Chironomus riparius 4th instar larvae previously exposed to microplastics of Polyurethane (PU-MPs). For this purpose, two investigations were carried out. In the first investigation, it was assessed the effects of PU-MPs (concentrations up to 750 mg/Kg) on C. riparius life history traits (larval size, emergence); and, when exposed to a no-observed-effect-concentration (NOEC), larvae behaviour (locomotion) and biochemical responses (oxidative damage, aerobic energy production, and energy reserves). Exposure to PU-MPs did not affect larval size nor imagoes emergence. Notwithstanding, contaminated larvae revealed higher aerobic metabolism underlined by a higher locomotion activity (curling/uncurling) but without compromising energy reserves. In the second experiment, it was assessed the effects of the consumption of contaminated alive prey (exposed to PU-MP NOEC) in a 3h feeding assay on planarians' behaviour (nr of consumed prey, locomotion), regeneration, and physiology (energy reserves, aerobic metabolism, oxidative damage). After the feeding test, planarians consumed 20% more contaminated prey with no changes in their locomotion. The consumption of contaminated prey delayed auricles' regeneration by one day, without resulting in oxidative damage, altered aerobic metabolism or energy reserves. These investigations bring new intel on the role of contaminated prey on planarians' behaviour and physiology in microplastic research.Os microplásticos (1 μm a 1 mm) tornaram-se um tópico preocupante na comunidade científica quando associados a uma má gestão de resíduos plásticos. Embora a presença e abundância destas partículas em ambientes de água doce tenha sido previamente reportada, ainda há um número insuficiente de estudos que fornecem informações sobre a potencial transferência trófica e as suas consequências nos organismos bentónicos de água doce. Este trabalho visou avaliar as respostas comportamentais e fisiológicas da planaria Girardia tigrina após o consumo de presas contaminadas - Chironomus riparius 4th instar larvae previamente expostas a microplásticos de Poliuretano (PU-MPs). Para este efeito, foram efetuadas duas investigações. Na primeira investigação, foram avaliados os efeitos dos PU-MPs (concentrações até 750 mg/Kg) sobre os parâmetros da história de vida de C. riparius (tamanho larvar, emergência); e, quando expostos a uma concentração sem efeitos observados (NOEC), comportamento larvar (locomoção) e respostas bioquímicas (danos oxidativos, produção de energia aeróbica, e reservas energéticas). A exposição a PU-MPs não afetou o tamanho das larvas nem a emergência dos adultos. Não obstante, as larvas contaminadas revelaram um metabolismo aeróbio mais elevado sublinhado por uma maior atividade de locomoção (N.º de contrações corporais) mas sem comprometer as reservas energéticas. Na segunda experiência, foram avaliados os efeitos do consumo de presas vivas contaminadas (expostas a PU-MP NOEC) num ensaio de alimentação de 3h sobre o comportamento das planárias (nº de presas consumidas, locomoção), regeneração, e fisiologia (reservas energéticas, metabolismo aeróbio, danos oxidativos). Após o ensaio de alimentação, as planárias consumiram mais 20% de presas contaminadas, sem alterações na sua locomoção. O consumo de presas contaminadas atrasou a regeneração das aurículas num dia, sem resultar em danos oxidativos, alteração do metabolismo aeróbico ou das reservas energéticas. Estas investigações trazem novas informações sobre o papel das presas contaminadas no comportamento e fisiologia das planárias na investigação de microplásticos.Mestrado em Eco-toxicologia e Análise de Risc