EFEITOS DOS ÓLEOS ESSENCIAIS DE ALECRIM (Rosmarinus officinalis) E PETITGRAIN (Citrus aurantium L.) EM MODELOS COMPORTAMENTAIS DE ATIVIDADE MOTORA, DEPRESSÃO, ANSIEDADE E APRENDIZADO EM RATOS

Apesar de várias evidências do potencial terapêutico dos óleos essenciais em diversas patologias, inclusive em transtornos mentais, os estudos científicos que comprovam esse potencial ainda são escassos. O objetivo deste trabalho foi investigar e comparar de forma sistemática os efeitos dos óleos essenciais de alecrim (Rosmarinus officinalis) e petitgrain (Citrus aurantium L.) em modelos animais com ratos nos seguintes parâmetros: atividade motora, depressão, ansiedade e aprendizado. Método: foram utilizados 297 ratos em todo o estudo, sendo: 54 no piloto 1; 66 no piloto 2; 36 no campo aberto; 36 na esquiva discriminativa; 36 no teste de enterrar esferas; 33 na natação forçada e 36 no experimento de aprendizagem. Os principais resultados revelaram que: ratos tratados com 100mg/kg (i.p.) de óleo essencial de alecrim não apresentaram diferença na atividade motora avaliada em campo aberto (p=0.213 teste de Mann-Whitney), tampouco na aprendizagem da resposta de pressão à barra em caixa de Skinner (p=0.098 teste de Mann-Whitney), comparados aos ratos controles que receberam salina 0,9% (1 mL/kg), porém esse mesmo tratamento foi efetivo em modelos de depressão (p=0.006 teste de Mann-Whitney) e ansiedade (teste de esconder esferas - p=0.003 ANOVA). No que diz respeito ao óleo essencial de petitgrain administrado em ratos na dose de 30mg/kg (i.p.), não observou-se diferença na atividade motora (p=0.795 teste de Mann-Whitney), contudo obteve-se efeito ansiolítico (teste de esconder esferas - p=0.028 ANOVA) e antidepressivo (p=0.001 teste de Mann-Whitney) em relação ao controle. Ademais, o óleo de petitgrain proporcionou uma melhora na aprendizagem (p=0.002 teste de Mann-Whitney) se comparado com os animais do grupo controle e os animais tratados com alecrim. Dessa forma podemos concluir que ambos os óleos estudados (alecrim e petitgrain) apresentaram atividades ansiolítica e antidepressiva nos testes realizados e apenas o óleo de petitgrain produziu efeitos na aprendizagem dos animais.

Melissa Romigh carrying wood to construct boardwalk in mangrove forest, Shark River Slough

This material is based upon work supported by the National Science Foundation through the Florida Coastal Everglades Long-Term Ecological Research program under Cooperative Agreements #DBI-0620409 and #DEB-9910514. This image is made available for non-commercial or educational use only.

Caractérisation moléculaire de la biodiversité des Fusarium associés à la fusariose de l'asperge (Asparagus officinalis L.) au Québec

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Caractérisation moléculaire de la biodiversité des Fusarium associés à la fusariose de l'asperge (Asparagus officinalis L.) au Québec

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Left to right: Dan Bond and Melissa Romigh. Setting up a flume panel to measure nutrient / sediment fluxed in a mangrove forest at SRS-6, Shark River Slough

This material is based upon work supported by the National Science Foundation through the Florida Coastal Everglades Long-Term Ecological Research program under Cooperative Agreements #DBI-0620409 and #DEB-9910514. This image is made available for non-commercial or educational use only.

Left to right: Gustavo Rubio and Melissa Romigh measuring diameter at breast height (DBH) of mangrove trees at SRS-6, Shark River Slough

This material is based upon work supported by the National Science Foundation through the Florida Coastal Everglades Long-Term Ecological Research program under Cooperative Agreements #DBI-0620409 and #DEB-9910514. This image is made available for non-commercial or educational use only.

Seawater carbonate chemistry and biological parameters of Sepia officinalis during experiments, 2009

Low pO2 values have been measured in the perivitelline fluids (PVF) of marine animal eggs on several occasions, especially towards the end of development, when embryonic oxygen consumption is at its peak and the egg case acts as a massive barrier to diffusion. Several authors have therefore suggested that oxygen availability is the key factor leading to hatching. However, there have been no measurements of PVF pCO2 so far. This is surprising, as elevated pCO2 could also constitute a major abiotic stressor for the developing embryo. As a first attempt to fill this gap in knowledge, we measured pO2, pCO2 and pH in the PVF of late cephalopod (Sepia officinalis) eggs. We found linear relationships between embryo wet mass and pO2, pCO2 and pH. pO2 declined from >12 kPa to less than 5 kPa, while pCO2 increased from 0.13 to 0.41 kPa. In the absence of active accumulation of bicarbonate in the PVF, pH decreased from 7.7 to 7.2. Our study supports the idea that oxygen becomes limiting in cephalopod eggs towards the end of development; however, pCO2 and pH shift to levels that have caused significant physiological disturbances in other marine ectothermic animals. Future research needs to address the physiological adaptations that enable the embryo to cope with the adverse abiotic conditions in their egg environment.

Seawater carbonate chemistry, mass, length and gene expression of Sepia officinalis during experiments, 2011

The specific transporters involved in maintenance of blood pH homeostasis in cephalopod molluscs have not been identified to date. Using in situ hybridization and immuno histochemical methods, we demonstrate that Na+/K+-ATPase (soNKA), a V-type H+-ATPase (soV-HA), and Na+/HCO3- cotransporter (soNBC) are co-localized in NKA-rich cells in the gills of Sepia officinalis. mRNA expression patterns of these transporters and selected metabolic genes were examined in response to moderately elevated seawater pCO2 (0.16 and 0.35 kPa) over a time-course of six weeks in different ontogenetic stages. The applied CO2 concentrations are relevant for ocean acidification scenarios projected for the coming decades. We determined strong expression changes in late stage embryos and hatchlings, with one to three log2-fold reductions in soNKA, soNBCe, socCAII and COX. In contrast, no hypercapnia induced changes in mRNA expression were observed in juveniles during both short- and long-term exposure. However a transiently increased demand of ion regulatory demand was evident during the initial acclimation reaction to elevated seawater pCO2. Gill Na+/K+-ATPase activity and protein concentration were increased by approximately 15% in during short (2-11 day), but not long term (42 day) exposure. Our findings support the hypothesis that the energy budget of adult cephalopods is not significantly compromised during long-term exposure to moderate environmental hypercapnia. However, the down regulation of ion-regulatory and metabolic genes in late stage embryos, taken together with a significant reduction in somatic growth, indicates that cephalopod early life stages are challenged by elevated seawater pCO2.

Seawater carbonate chemistry and trace element accumulation during experiments with common cuttlefish, Sepia officinalis, 2009

Cephalopods play a key role in many marine trophic networks and constitute alternative fisheries resources, especially given the ongoing decline in finfish stocks. Along the European coast, the eggs of the cuttlefish Sepia officinalis are characterized by an increasing permeability of the eggshell during development, which leads to selective accumulation of essential and non-essential elements in the embryo. Temperature and pH are two critical factors that affect the metabolism of marine organisms in the coastal shallow waters. In this study, we investigated the effects of pH and temperature through a crossed (3?2; pH 8.1 (pCO2, 400 ppm), 7.85 (900 ppm) and 7.6 (1400 ppm) at 16 and 19°C, respectively) laboratory experiment. Seawater pH showed a strong effect on the egg weight and non-significant impact on the weight of hatchlings at the end of development implying an egg swelling process and embryo growth disturbances. The lower the seawater pH, the more 110 mAg was accumulated in the tissues of hatchlings. The 109Cd concentration factor (CF) decreased with decreasing pH and 65Zn CF reached maximal values pH 7.85, independently of temperature. Our results suggest that pH and temperature affected both the permeability properties of the eggshell and embryonic metabolism. To the best of our knowledge, this is one of the first studies on the consequences of ocean acidification and ocean warming on metal uptake in marine organisms, and our results indicate the need to further evaluate the likely ecotoxicological impact of the global change on the early-life stages of the cuttlefish.

Seawater carbonate chemistry and biological processes of Sepia officinalis during experiments, 2010

Acidification of ocean surface waters by anthropogenic carbon dioxide (CO2) emissions is a currently developing scenario that warrants a broadening of research foci in the study of acid-base physiology. Recent studies working with environmentally relevant CO2 levels, indicate that some echinoderms and molluscs reduce metabolic rates, soft tissue growth and calcification during hypercapnic exposure. In contrast to all prior invertebrate species studied so far, growth trials with the cuttlefish Sepia officinalis found no indication of reduced growth or calcification performance during long-term exposure to 0.6 kPa CO2. It is hypothesized that the differing sensitivities to elevated seawater pCO2 could be explained by taxa specific differences in acid-base regulatory capacity. In this study, we examined the acid-base regulatory ability of S. officinalis in vivo, using a specially modified cannulation technique as well as 31P NMR spectroscopy. During acute exposure to 0.6 kPa CO2, S. officinalis rapidly increased its blood [HCO3] to 10.4 mM through active ion-transport processes, and partially compensated the hypercapnia induced respiratory acidosis. A minor decrease in intracellular pH (pHi) and stable intracellular phosphagen levels indicated efficient pHi regulation. We conclude that S. officinalis is not only an efficient acid-base regulator, but is also able to do so without disturbing metabolic equilibria in characteristic tissues or compromising aerobic capacities. The cuttlefish did not exhibit acute intolerance to hypercapnia that has been hypothesized for more active cephalopod species (squid). Even though blood pH (pHe) remained 0.18 pH units below control values, arterial O2 saturation was not compromised in S. officinalis because of the comparatively lower pH sensitivity of oxygen binding to its blood pigment. This raises questions concerning the potentially broad range of sensitivity to changes in acid-base status amongst invertebrates, as well as to the underlying mechanistic origins. Further studies are needed to better characterize the connection between acid-base status and animal fitness in various marine species.

Seawater carbonate chemistry and biological processes during experiments with common cuttlefish Sepia officinalis, 2010

Ocean acidification and associated changes in seawater carbonate chemistry negatively influence calcification processes and depress metabolism in many calcifying marine invertebrates. We present data on the cephalopod mollusc Sepia officinalis, an invertebrate that is capable of not only maintaining calcification, but also growth rates and metabolism when exposed to elevated partial pressures of carbon dioxide (pCO2). During a 6 wk period, juvenile S. officinalis maintained calcification under ~4000 and ~6000 ppm CO2, and grew at the same rate with the same gross growth efficiency as did control animals. They gained approximately 4% body mass daily and increased the mass of their calcified cuttlebone by over 500%. We conclude that active cephalopods possess a certain level of pre-adaptation to long-term increments in carbon dioxide levels. Our general understanding of the mechanistic processes that limit calcification must improve before we can begin to predict what effects future ocean acidification will have on calcifying marine invertebrates.

Seawater carbonate chemistry and biological processes during experiments with common cuttlefish Sepia officinalis, 2008

Ocean acidification and associated changes in seawater carbonate chemistry negatively influence calcification processes and depress metabolism in many calcifying marine invertebrates. We present data on the cephalopod mollusc Sepia officinalis, an invertebrate that is capable of not only maintaining calcification, but also growth rates and metabolism when exposed to elevated partial pressures of carbon dioxide (pCO2). During a 6 wk period, juvenile S. officinalis maintained calcification under ~4000 and ~6000 ppm CO2, and grew at the same rate with the same gross growth efficiency as did control animals. They gained approximately 4% body mass daily and increased the mass of their calcified cuttlebone by over 500%. We conclude that active cephalopods possess a certain level of pre-adaptation to long-term increments in carbon dioxide levels. Our general understanding of the mechanistic processes that limit calcification must improve before we can begin to predict what effects future ocean acidification will have on calcifying marine invertebrates.

Phenolic composition of four sage species: salvia farinacea, salvia mexico, salvia greggii and salvia officinalis

Salvia species are used worldwide for medicine purposes. In general, these medicinal plants have high amounts of flavonoids and phenolic acids, that are thought to be closely related to their health properties [1,2]. In this work, the aerial parts of Salvia farinacea, Salvia mexico, Salvia greggii and Salvia officinalis were extracted with hot water [3]. Extracts were evaluated for their total phenolic content by an adaptation of the Folin-Ciocalteu method and further analysed by high performance liquid chromatography associated with electrospray mass spectrometry (HPLC-DAD-ESI-MSn) in the negative ion mode [4], in order to identify their individual phenolic constituents. The aqueous extracts of S. farinacea, S. mexico, S. officinalis and S. greggii contained, respectively, 106±13, 159±38, 175±46 and 136±1 μg GAE/mg of total phenolics. These four species were characterized by a clear prevalence of caffeic acid derivatives, in particular of rosmarinic acid (MW 360), that is generally the most abundant phenolic compound in Salvia species [2,3]. In addition, S. mexico and S. officinalis contained moderate amounts of salvianolic acid B (MW 718). Among these two, S. mexico was richer in O-caffeoylquinic acid (MW 354), while the latter presented high amounts of salvianolic acid K (MW 556) and moderate amounts of its structural isomer. All the extracts were enriched in flavones: S. farinacea and S. officinalis contained high amounts of luteolin-O-glucuronide while S. mexico contained luteolin-C-glucoside with respective characteristic mass spectrometry fragmentation pattern m/z at 461→285 and m/z at 447→357, 327. Similarly, S. greggii extract presented high content of luteolin-7-O-glucoside ([M-H]− at m/z 447→ 285) and luteolin-C-glucoside and moderate quantities of apigenin-C-hexoside ([M-H]− at m/z 431→341, 311). Further studies are being undertaken in order to understand the contribution of these phenolic constituents in the biological activities of Salvia plants.

Phenolic composition of four sage species: salvia farinacea, salvia mexico, salvia greggii and salvia officinalis

Salvia species are used worldwide for medicine purposes. In general, these medicinal plants have high amounts of flavonoids and phenolic acids, that are thought to be closely related to their health properties [1,2]. In this work, the aerial parts of Salvia farinacea, Salvia mexico, Salvia greggii and Salvia officinalis were extracted with hot water [3]. Extracts were evaluated for their total phenolic content by an adaptation of the Folin-Ciocalteu method and further analysed by high performance liquid chromatography associated with electrospray mass spectrometry (HPLC-DAD-ESI-MSn) in the negative ion mode [4], in order to identify their individual phenolic constituents. The aqueous extracts of S. farinacea, S. mexico, S. officinalis and S. greggii contained, respectively, 106±13, 159±38, 175±46 and 136±1 μg GAE/mg of total phenolics. These four species were characterized by a clear prevalence of caffeic acid derivatives, in particular of rosmarinic acid (MW 360), that is generally the most abundant phenolic compound in Salvia species [2,3]. In addition, S. mexico and S. officinalis contained moderate amounts of salvianolic acid B (MW 718). Among these two, S. mexico was richer in O-caffeoylquinic acid (MW 354), while the latter presented high amounts of salvianolic acid K (MW 556) and moderate amounts of its structural isomer. All the extracts were enriched in flavones: S. farinacea and S. officinalis contained high amounts of luteolin-O-glucuronide while S. mexico contained luteolin-C-glucoside with respective characteristic mass spectrometry fragmentation pattern m/z at 461→285 and m/z at 447→357, 327. Similarly, S. greggii extract presented high content of luteolin-7-O-glucoside ([M-H]− at m/z 447→ 285) and luteolin-C-glucoside and moderate quantities of apigenin-C-hexoside ([M-H]− at m/z 431→341, 311). Further studies are being undertaken in order to understand the contribution of these phenolic constituents in the biological activities of Salvia plants.

Avaliação de micronutrientes e sua influência no metabolismo secundário de Bidens pilosa e Salvia officinalis, plantas usadas no tratamento de diabetes

Dissertação (Mestrado em Tecnologia Nuclear)