The aim of these studies was to identify circulating miRNAs that can be used as biomarkers in patients with paracetamol-induced liver injury. Whether the miRNAs discovered in humans could be back-translated to zebrafish with the aim of developing a liver toxicity model to replace rodent use was also investigated. First, the miRNA signature of DILI induced by paracetamol was defined. Plasma miRNAs were quantified in paracetamol overdose patients. A signature of 16 miRNAs was discovered that best separated patients with liver injury from those without liver injury. This signature was tested in a second cohort and resulted in the detection of paracetamol-induced liver injury with high specificity and sensitivity. At first presentation to hospital miR-122-5p was the most sensitive single miRNA and superior to ALT activity in predicting liver injury. In order to further qualify miR-122-5p, three detailed studies relevant to possible clinical scenarios were performed. The effect of acute alcohol ingestion (commonly co-ingested with paracetamol overdose) on circulating concentrations of miR-122-5p in healthy volunteers was investigated. Alcohol ingestion induced a small, non-clinically relevant, increase in miR-122-5p. The effect of chronic kidney disease (CKD) and haemodialysis (HD) on circulating miR-122-5p concentrations was explored because kidney dysfunction has been associated with a reduction in the concentration of circulating miRNAs. HD patients had lower concentrations of miR- 122-5p compared to healthy volunteers and CKD patients. To facilitate miRNA measurement outwith hospitals, miR-122-5p was measured in a blood drop from a finger prick. miR-122-5p was readily measurable in finger prick samples and concentrations were significantly higher in the blood drop from DILI patients compared with healthy volunteers. To complement miR-122-5p as a marker of toxicity, circulating paracetamol metabolites were measured in plasma samples from paracetamol overdose patients. A higher percentage of circulating metabolites formed by cytochrome P450 enzymes were present in patients with liver injury and these metabolites were superior to both ALT and paracetamol concentration with regard to early patient stratification. To reduce need for rodent studies, miRNAs were back-translated into zebrafish. In order to study circulating miR-122-5p in adult zebrafish, a bloodletting method by collecting blood retro-orbitally was developed. After studying different dosing regimens of paracetamol in adult and larvae zebrafish the model was determined to be too variable with regard to liver injury. A new drug, triptolide, originating from traditional Chinese medicine and responsible for DILI in China, was tested as an alternative model for drug-induced liver injury in zebrafish larvae. miRNA-122-5p decreased in zebrafish larvae after triptolide treatment and triptolide-induced liver injury could be tracked by fluorescent microscopy. Selective plane illumination microscopy was able to track the decrease in liver volume during triptolide exposure. In order to identify the toxic pathways involved in triptolide-induced liver injury, RNA-sequencing was performed. This identified KEGG pathways including ribosome, spliceosome and notch signalling as pathways affected by triptolide. In summary, miRNAs can be used as highly sensitive biomarkers to detect acute liver injury in patients and zebrafish. Zebrafish may represent an alternative model species to study DILI, further work is needed.

Delta-like homologue 1 (Dlk1) and Dlk2 encode vertebrate-specific transmembrane proteins belonging to the Jagged/Delta/Serrate family of Notch ligands. Murine Dlk1 is widely expressed during embryonic development and targeted deletion results in defects in numerous developmental processes, such as adipogenesis, haematopoiesis, neurogenesis and skeletal muscle formation. However, the mechanisms by which DLK1 regulates these processes remains unclear. The purpose of this project is to examine the function of these genes using zebrafish as an in vivo model, allowing insight to the ancestral functions of these genes. We have strong evolutionary evidence that dlk2 is the ancestral version of the gene from which dlk1 is derived; therefore, the thesis focuses primarily on the role of dlk2 in the zebrafish system. I initially examine the expression of zebrafish dlk1 and dlk2 during embryonic development and in the adult brain, determining similarities and differences between mouse and zebrafish. In particular, dlk1 and dlk2 in the fish exhibit a pattern that is more reminiscent of Dlk2 in the mouse. This developmental expression pattern is essential for the interpretation of the modulation of Dlk2 in later chapters, and is aided by the generation of a mammalian Dlk2 antibody that cross-reacts with zebrafish. We obtained a dlk2 mutant and used this line to examine the role of the DLK2 protein in development and in the adult brain. I demonstrate that, in the absence of DLK2, a population of neural precursor cells appear to over-proliferate early in zebrafish development. Later, by larval stages, these cells are absent, suggesting a premature activation and subsequent depletion of the progenitor cell pool in the mutant, reminiscent of the Dlk1 mutant in mouse. Associated with this phenotype are larval behavioral defects in motor response. In this thesis, it will be shown that in the adult dlk2 mutant zebrafish, the radial glial cell population in the telencephalon is completely depleted. These radial glial cells are thought to be responsible for adult neural regeneration in zebrafish, and our characterization of a mutant completely lacking this cell population provides a rich model to further examine and understand the functions of this well-studied but poorly understood cell population. These findings have both functional and evolutionary implications for the relative roles of these two vertebrate specific atypical Notch ligands.

Cryopreservation of fish germ cells has important applications in aquaculture, conservation of endangered species and human genomic studies. Although investigations on cryopreservation of fish sperm and embryos have been carried out extensively, cryopreservation of fish oocytes has not been studied systematically. The objective of the present study was to develop successful cryopreservation protocol for zebrafish oocytes at temperature of liquid nitrogen (-196°C), or if unachieved, to investigate the limiting factors associated with fish oocytes cryopreservation. In this study, the effects of cryoprotectants exposure and enzymatic treatments on oocytes survival were studied, and new viability tests for zebrafish oocytes were developed. The effects of controlled slow cooling with different cryoprotective agents, in different freezing media and at different cooling rates on cryosurvival of zebrafish (D. rerio) oocytes were investigated. Cryomicroscopic observations on zebrafish oocytes were also carried out. Three reliable vital tests -trypan blue (TB) staining, ATP assay, and in vitro maturation followed by germinal vesicle breakdown observation (GVBD) were found suitable for assessment of oocytes viability. Vitellogenesis (stage III) was found to be the optimal developmental stage for cryopreservation. Methanol was found to be the best CPA for zebrafish oocytes. Combination of 4M methanol and 0.2M glucose in potassium chloride (KCI) buffer was found to be the optimal cryoprotective solution. Controlled slow cooling at 0.3°C/min rate, combined with seeding at -12.5°C and plunge to liquid nitrogen (LN) at-40°C were found to be the optimal conditions for cryopreservation of stage III oocytes. However, even with the optimal protocol, TB-assessed viability, Le. the ratio of oocytes with intact plasma membrane after cooling to -196°C was 19.6±8%. Furthermore, GVBD experiments showed that none of the cryopreserved oocytes can be matured in vitro, and their ATP levels were decreased dramatically, indicating that successful cryopreservation of fish oocytes at liquid nitrogen temperature still remains elusive. Cryomicroscopic observations demonstrated, that the damages of oocytes are associated with intracellular ice formation (lIF). IIF occurred simultaneously with extracellular ice formation (ElF) in nearly 100% of the cases, and formation of lethal hexagonal type of ice was observed. This study was the first systematic attempt to cryopreserve fish oocytes at liquid nitrogen temperature. The results provided will undoubtedly assist successful protocol design for cryopreservation of fish oocytes in the future.

xvii, 125 p. : ill. (some col.) A print copy of this thesis is available through the UO Libraries. Search the library catalog for the location and call number. / MicroRNAs are small regulatory RNAs that control various developmental and physiological processes in animals and plants. To study the involvement of microRNAs in skeletal development, I manipulated the expression of miR-140, which is strongly expressed in the developing skeleton, and miR-196, which is located among the body patterning Hox cluster genes. I found that miR-140 regulates zebrafish palate formation by interfering with neural crest cell migration through the inhibition of the expression of the platelet derived growth factor receptor alpha ( pdgfra ) gene. I also found that miR196 regulates zebrafish pectoral fin initiation by regulating the expression of the retinoic acid receptor alpha b ( rarab ) gene and that miR-196 is involved in the patterning of zebrafish pharyngeal arches and vertebrae. These results illuminate previously unknown regulatory mechanisms of skeletal development. I also reviewed current knowledge concerning microRNAs in skeletal development and evolution and discussed potential relationships between microRNAs and skeletal disease. This dissertation includes previously published and unpublished coauthored material. / Committee in charge: Judith Eisen, Chairperson, Biology; John Postlethwait, Advisor, Biology; Charles Kimmel, Member, Biology; William Cresko, Member, Biology; J. Andrew Berglund, Outside Member, Chemistry

Angiogenesis is the process of new blood vessel sprouting from pre-existing vessels and is responsible for generating the majority of nascent vessels during development, tissue regeneration and disease. During angiogenesis, sprouting endothelial cells (ECs) are organised into leading 'tip' cells (TCs) and trailing 'stalk' cells (SCs). This hierarchal organisation of TCs and SCs is essential for the coordinated collective migration of ECs during sprouting. However, the precise mechanisms that define TC verses SC behaviour and identity remains uncertain. Transcriptomic analysis of sprouting vessels in zebrafish embryos led to the identification of a novel TC-associated gene, transmembrane 4 L six family member 1 (tm4sf1). We find that tm4sf1 expression is tightly spatiotemporally restricted to migrating TCs during intersegmental vessel (ISV) sprouting in zebrafish. Furthermore, TC tm4sf1 expression is controlled by the vascular endothelial growth factor receptor (Vegfr) - Notch signalling axis. Morpholino oligonucleotide (MO)-mediated knockdown of tm4sf1 reveals a subtle delay in ISV sprouting upon loss of tm4sf1 expression. Moreover, using multiplexed, real-time imaging approaches and in-depth analysis of TC and SC behaviours at single cell resolution, we reveal that the delay in ISV sprouting is specifically due to reduced TC motility. Furthermore, we find that tm4sf1 functions to induce TC motility in the leading daughter cell following TC mitosis, to rapidly re-establish post-mitotic TC behaviour. Generation of tm4sf1 mutant zebrafish lines using both transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR) confirms that Tm4sf1 modulates TC behaviour. Additionally, mechanistic studies in human ECs reveal that tm4sf1 regulates VEGFR-mediated signalling upon VEGF-stimulation, which subsequently controls cell migration and expression of the TC determinants, DLL4 and VEGFR2. Hence, our results suggest that tm4sf1 is a novel modulator of the TC-SC hierarchy and collective EC migration during ISV sprouting. Overall, these findings have potential therapeutic implications since tm4sf1 may be a promising target for the manipulation of pathological angiogenesis in disease.

Purely diffusive O2 transport typically is insufficient to sustain aerobic metabolism in most multicellular organisms. In small animals, however, a high surface-to-volume ratio may allow passive diffusion alone to supply sufficient O2 transfer. The purpose of this thesis was to explore the impacts of internal convection on the exchange of respiratory gases in a small complex organism, the larval zebrafish (Danio rerio). Thus, I tested the hypothesis that internal convection is required for the normal transfer of the respiratory gases O2 and CO2 and maintenance of resting aerobic metabolic rate. Use of morpholino knockdown of the VEGF-A and TNNT2 proteins allowed examination of two independent models lacking internal convection. Using micro-respirometry, I demonstrated that loss of internal convection reduces resting rates of O2 consumption and CO2 excretion in larvae at 4 days post fertilization. I also used the scanning micro-optrode technique to demonstrate that acute loss of internal convection resulted in reduced rates of cutaneous O2 flux, a trait that was reversed upon the restoration of internal convection. Finally, I demonstrated that in larval zebrafish, loss of internal convection resulted in decreased hypoxic performance and loss or severe reduction of the hypoxic cardiorespiratory responses. The results from these experiments showed that internal convection is i) required to maintain resting rates of respiratory gas transfer in the larval zebrafish, ii) important in facilitating the hypoxic cardiorespiratory responses in larval zebrafish and iii) augments O2 extraction capacity in the face of progressive hypoxia.

Submitted by HELENA MANNOCHIO RUSSO null (helenamrusso@gmail.com) on 2018-03-20T18:22:07Z No. of bitstreams: 1 Dissertação Helena M Russo.pdf: 9145473 bytes, checksum: 2f4fdbd123ab5e669c33fc38d6613f35 (MD5) / Approved for entry into archive by Ana Carolina Gonçalves Bet null (abet@iq.unesp.br) on 2018-03-23T18:49:14Z (GMT) No. of bitstreams: 1 russo_hm_me_araiq_int.pdf: 9117178 bytes, checksum: 84527f550234fa1c49bf57f9a53f5f71 (MD5) / Made available in DSpace on 2018-03-23T18:49:14Z (GMT). No. of bitstreams: 1 russo_hm_me_araiq_int.pdf: 9117178 bytes, checksum: 84527f550234fa1c49bf57f9a53f5f71 (MD5) Previous issue date: 2018-03-02 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / A espécie Tetrapterys mucronata (Malpighiaceae), uma das espécies ocasionalmente utilizados na Ayahuasca, despertou há algum tempo o interesse do grupo de pesquisa NuBBE para a realização do estudo químico e avaliação das atividades biológicas devido a sua utilização nos rituais religiosos conhecidos como “Santo Daime”. Este estudo revelou resultados interessantes nos ensaios preliminares voltados para a identificação de potenciais inibidores de acetilcolinesterase, e mostrou que a espécie T. mucronata tem alcaloides e compostos fenólicos como principais constituintes químicos. Estes resultados levaram ao estudo de outras espécies de Tetrapterys ainda sem estudo químico, incluindo espécies que pertenciam a esse gênero e recentemente foram reclassificados como Niedenzuella, como no caso da Niedenzuella multiglandulosa, espécie selecionada para esta pesquisa de mestrado. Esta espécie é conhecida devido aos efeitos tóxicos que causam a morte de rebanhos bovinos, e de outros mamíferos, em geral, causando prejuízos substanciais na balança comercial brasileira. Este trabalho tem como objetivo a busca de compostos tóxicos presentes nas folhas de Tetrapterys multiglandulosa. Essa espécie foi selecionada por serem relatados na literatura surtos de intoxicação do gado (em 2004 e 2005) em áreas altamente infestadas pela planta, causando morte de vacas e aborto de bezerros no estado do Mato Grosso do Sul. Estudos recentes encontrados na literatura indicam a presença de monofluoracetato (MF), uma substância conhecidamente tóxica, nessa espécie, mas ainda não foram realizados estudos fitoquímicos e de toxicidade para esta espécie e nem mesmo para o gênero Niedenzuella. A partir de folhas coletadas no próprio estado onde os surtos foram relatados, extratos foram preparados utilizando solventes de diferentes polaridades e enviados para ensaio de toxicidade empregando modelo com Zebrafish, cujo resultado mostrou que o extrato metanólico era o único tóxico. O foco do estudo voltou-se para esse extrato, sendo realizado um fracionamento utilizando a técnica de MPLC-UV e, em seguida, uma purificação mais acurada por HPLC em escala preparativa. Foram isoladas 19 substâncias das classes dos alcaloides, flavonoides glicosilados e não glicosilados, compostos glicosilados e 6 esteroides (denominados ecdisteroides), ainda não relatados para esse gênero, incluindo um ecdisteroide inédito (17), que foram identificadas e caracterizadas por UHPLC-TOF-HRMS e NMR. As substâncias isoladas foram identificadas como: trigonelina (1), triptofano (2), 4- hidroxicinamamida (3), ácido (Z)-p-cumárico 4-O-β-D-glicopiranosídeo (4), icarisida F2 (5), luteoforol (6), ácido (E)-4-hidroxicinâmico (7), ácido (Z)-4-hidroxicinâmico (8), integristerona A (9), epiecdisterona (11), kaempferol triglicosilado (12), ecdisterona (13), isorhamnetina triglicosilada (14), ácido cinâmico (16), multiglandisterona (17), calonisterona (18) e podecdisona B (19). O esteroide majoritário (13) foi testado novamente para toxicidade utilizando modelo de Zebrafish e se mostrou tóxico, também um dado inédito para a espécie em estudo. Além disso, as moléculas isoladas são bastante diferentes das já identificadas para os gêneros de Tetrapterys, o que corroboram para a diferenciação dessas espécies em dois gêneros distintos. Análises de NMR 19F e LC-MS foram realizadas para tentar identificar a presença de MF nos extratos e frações polares, no entanto, não foi possível. Os resultados obtidos corroboram para um maior entendimento de espécies tóxicas para gado presentes no Brasil, e apresentam dados complementares aos publicados recentemente. / Tetrapterys mucronata (Malpighiaceae) species, one of the species occasionally used in Ayahuasca, has become of interest of NuBBE research group to perform a chemical study for an evaluation of the biological activities due to its use in religious rituals known as “Santo Daime”. This study revealed interesting results in preliminary assays aiming to identify acetylcholinesterase potential inhibitors, and that this species has alkaloids and phenolic compounds as its major chemical compounds. These results led our research group to study other species from Tetrapterys genus without further chemical studies, including species that used to belong to this genus and that recently has changed to Niedenzuella genus, such as Niedenzuella multiglandulosa, the selected species for this master’s research. This species is known for causing toxic effects that may cause death in cattle and other mammals in general, causing substantial losses in Brazilian trade balance. The objective of this research is to search for toxic compounds in Niedenzuella multiglandulosa leaves. This species was chosen for having reports in the literature of outbreaks of cattle intoxication (in 2004 and 2005) in which the pasture was highly infested with it, causing death of cows and led to abortion of calves in Mato Grosso do Sul state. Recent studies found in the literature indicate the presence of monofluoracetate (MFA), a well-known toxic substance, in this species, but so far there were no phytochemical and toxicity assay study regarding this species, and not even for the genus Niedenzuella. From the leaves collected in the same state where there were the outbreaks were reported, extracts were prepared using different polarity solvents and sent to Zebrafish toxicity assay, whose result showed that the methanolic extract was the only toxic one. The focus of this study turned to this extract, performing a fractionation using MPLC-UV, and then a more accurate purification using preparative HPLC. 19 substances were isolated such as alkaloids, glycosylated and non- glycosylated flavonoids, glycosylated compounds and 6 steroids (also called ecdysteroids), including a new ecdysteroid (17) that were identified and elucidated by UHPLC-TOF-HRMS and NMR. The isolated substances were identified as: trigonelline (1), tryptophan (2), 4-hydroxycinnamamide (3), p- Coumaric acid 4-O-β-D-glucopyranoside (4), icariside F2 (5), luteoforol (6), (E)-4- Hydroxycinnamic acid (7), (Z)-4-Hydroxycinnamic acid (8), integristerone A (9), epiecdysterone (11), kaempferol triglucoside (12) ecdysterone (13), isorhamnetin triglycoside (14), cinnamic acid (16), multiglandysterone (17), calonysterone (18) e Podecdysone B (19). The major steroid (13) was tested again for toxicity using Zebrafish model and showed toxicity, also a new data for the species in study. Additionally, the isolated compounds are quite different from the isolated previously from Tetrapterys genus, which corroborate to the fact that these species belong to different genera. 19F NMR and LC-MS analysis were performed attempting to identify the presence of MFA in polar extracts and fractions, however it wasn’t possible to identify it. The obtained results corroborate to a better understanding of toxic / 152341/2015-2

Submitted by GUILHERME EDUARDO SASSI null (gui_sassi@hotmail.com) on 2016-07-07T23:52:04Z No. of bitstreams: 1 Impressão Final com Ficha.pdf: 1062820 bytes, checksum: d13b320c892331bf84bf85a95aa7b409 (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-07-12T12:46:39Z (GMT) No. of bitstreams: 1 sassi_ge_me_bot.pdf: 1062820 bytes, checksum: d13b320c892331bf84bf85a95aa7b409 (MD5) / Made available in DSpace on 2016-07-12T12:46:39Z (GMT). No. of bitstreams: 1 sassi_ge_me_bot.pdf: 1062820 bytes, checksum: d13b320c892331bf84bf85a95aa7b409 (MD5) Previous issue date: 2016-06-15 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O zebrafish, teleósteo de água doce, apresenta características biológicas e genéticas favoráveis para pesquisas biológicas, entretanto, faltam informações sobre sua exigência nutricional. Neste sentido este trabalho objetivou determinar sua exigência proteica. Utilizou-se 900 juvenis com 25,2±1,98 mg e 12,95 ± 1,91 mm, distribuídos em 30 aquários de 15 L de volume (30 peixes/aquário) num delineamento inteiramente casualizado com cinco tratamentos e seis repetições. As dietas semipurificadas foram formuladas para conter 27,6; 33,2; 38,2; 43,8 e 49,2% de proteína bruta (PB). Após 62 dias, constatou-se que o teor de 27.6% PB foi mais efetivo (p<0.01) para a taxa de eficiência proteica. Entretanto, não houve diferença entre tratamentos para peso, comprimento, taxa de eficiência alimentar e de crescimento especifico, consumo de ração e sobrevivência. Em relação a utilização dos nutrientes e composição corporal, a dieta contendo 38,2% de PB proporcionou melhores respostas (p<0.01) para retenção e taxa de retenção de nitrogênio, retenção de energia e quantidade de energia corporal. Pode-se concluir que, dieta contendo 27.6% PB foi adequada para obtenção da melhor taxa de eficiência proteica e desempenho zootécnico e 38,2% PB para utilização dos nutrientes e energia pelo zebrafish. / The zebrafish, teleost freshwater, shows biological and genetic characteristics tha are propitious for biological research . However, there is a lack of information about their nutritional requirements, considering this the purpose of this study is to determine its protein requirement. Were used 900 fingerlings with 25,2±1,98 mg distributed into 30 fish tanks of 15 liters of capacity (30 fishes/fish tank) in a completely randomized with five treatment and six repetitions. The Semipurified diets were formulated to include 27.6; 33.2; 38.2; 43.8; 49,2% of crude protein (CP). After 62 days, it was found that the percentage of 27.6% CP were more efective (p<0.01) to protein efficiency ratio. Although there was no difference between treatments for weight, length, feed efficiency rate and specific growth, feed intake and survival. In relation the use of nutrients and body composition, diet containing 38.2% crude protein provided better responses (p<0,01) for retention and nitrogen retention rate, energy retention and amount of body energy. We can be concluded that a diet containing 27.6% CP was adequate to obtain the best protein efficiency ratio and growth performance and 38.2% CP for use of nutrients and energy for the zebrafish.

Submitted by Mario BC (mario@bc.ufrpe.br) on 2018-06-18T13:53:36Z No. of bitstreams: 1 Tiago Queiroz da Mota Bittencourt.pdf: 3519794 bytes, checksum: 6c8d40c94975ee0c835f086dd5e829ec (MD5) / Made available in DSpace on 2018-06-18T13:54:56Z (GMT). No. of bitstreams: 1 Tiago Queiroz da Mota Bittencourt.pdf: 3519794 bytes, checksum: 6c8d40c94975ee0c835f086dd5e829ec (MD5) Previous issue date: 2017-02-24 / The toxic effects of sodium metavanadate (MV), vanadium pentoxide (PV) and oxovanadium sulfate (SV) on embryos and adults of zebrafish (D. rerio) were evaluated. Embryos were exposed to concentrations of 10-1000 μg/mL for evaluation of 96-h LC50 and their teratogenic effects. Adults were exposed to 20 μg/mL to evaluate behavioral changes related to vanadium compounds exposure and daily survival. Embryos showed a 96-h LC50 of 74.14 for PV and 100% mortality at the concentrations of 400-1000 μg/mL of the three compounds. The most significant teratogenic effects were pericardial edema and yolk sac (p < 0.05). The adults exposed to the vanadium compounds presented higher opercular beating and darkening of the gill arches. Floating and Resting behaviors in exposed adults were significant (p < 0.05), as well as the behavior of air breathing behavior. It can be concluded that in tests with the embryos the vanadium pentoxide is the most toxic compound and that pericardial edema and yolk sac were the most significant teratogenic effects and that the exposure of adults to vanadium compounds promoted an increase in the frequency of behaviors Inactive and stress response. / Foram avaliados os efeitos tóxicos do metavanadato de sódio (MV), pentóxido de vanádio (PV) e sulfato de oxovanádio (SV) em embriões e adultos de zebrafish (D. rerio). Os embriões foram expostos a concentrações de 10-1000 μg/mL para avaliação da CL50 96 h e seus efeitos teratogênicos. Os adultos foram expostos a 20 μg/mL para avaliar alterações comportamentais relacionadas à exposição dos compostos de vanádio e sobrevivência diária. Os embriões apresentaram uma CL50 96 h de 74,14 para o PV e 100% de mortalidade nas concentrações de 400-1000 μg/mL dos três compostos. Os efeitos teratogênicos mais significativos foram edemas de pericárdio e saco vitelínico (p < 0,05). Os adultos expostos aos compostos de vanádio apresentaram maior batimento opercular e escurecimento dos arcos branquiais. A exibição dos comportamentos Flutuar e Descansar nos adultos expostos foram significativos (p < 0,05), como também a exibição do comportamento Respiração aérea. Pode-se concluir que nos testes com os embriões o pentóxido de vanádio é o composto mais tóxico e que edemas de pericárdio e saco vitelínico foram os efeitos teratogênicos mais significativo e que a exposição dos adultos aos compostos de vanádio promoveram um aumento na frequência dos comportamentos inativos e de resposta ao estresse.

To date, limited research has been conducted on the effects of cytotoxic drugs on aquatic species. Three of these species, a single celled plant, an invertebrate and a fish, were exposed to six cytotoxic drugs, both singly and as simple mixtures. A range of endpoints were assessed, including growth inhibition in algae, immobilisation in daphnia and the impact on organ size and the expression of the DNA damage repair genes RAD51 and p53 in zebrafish. No effects at environmentally relevant concentrations were observed in any of the three species.