The effects of temperature on nickel tocicity in goldfish (carassius auratus L.)

Although it is widely assumed that temperature affects pollutant toxicity, few studies have actually investigated this relationship. Moreover, such research as has been done has involved constant temperatures; circumstances which are rarely, if ever, actually experienced by north temperate, littoral zone cyprinid species. To investigate the effects of temperature regime on nickel toxicity in goldfish (Carassius auratus L.), 96- and 240-h LCSO values for the heavy metal pollutant, nickel (NiCI2.6H20), were initially determined at 2DoC (22.8 mg/L and 14.7 mg/L in artificially softened water). Constant temperature bioassays at 10°C, 20°C and 30°C were conducted at each of 0, 240-h and 96-h LCSO nickel concentrations for 240 hours. In order to determine the effects of temperature variation during nickel exposure it was imperative that the effects of a single temperature change be investigated before addressing more complex regimes. Single temperature changes of + 10°C or -10°C were imposed at rates of 2°C/h following exposures of between 24 hand 216 h. The effects of a single temperature change on mortality, and duration of toxicant exposure at high and low temperatures were evaluated. The effects of fluctuating temperatures during exposure were investigated through two regimes. The first set of bioassays imposed a sinewave diurnal cycle temperature (20.±.1DOC) throughout the 10 day exposure to 240-h LeSO Ni. The second set of investigations approximated cyprinid movement through the littoral zone by imposing directionally random temperature changes (±2°C at 2-h intervals), between extremes of 10° and 30°C, at 240-h LC50 Ni. Body size (i.e., total length, fork length, and weight) and exposure time were recorded for all fish mortalities. Cumulative mortality curves under constant temperature regimes indicated significantly higher mortality as temperature and nickel concentration were increased. At 1DOC no significant differences in mortality curves were evident in relation to low and high nickel test concentrations (Le., 16 mg/L and 20 mg/L). However at 20°C and 30°C significantly higher mortality was experienced in animals exposed to 20 mg/L Ni. Mortality at constant 10°C was significantly lower than at 30°C with 16 mg/L and was significantly loWer than each of 2DoC and 39°C tanks at 20 mg/L Ni exposure. A single temperature shift from 20°C to 1DoC resulted in a significant decrease in mortality rate and conversely, a single temperature shift from 20°C to 30°C resulted in a significant increase in mortality rate. Rates of mortality recorded during these single temperature shift assays were significantly different from mortality rates obtained under constant temperature assay conditions. Increased Ni exposure duration at higher temperatures resulted in highest mortality. Diurnally cycling temperature bioassays produced cumulative mortality curves approximating constant 20°C curves, with increased mortality evident after peaks in the temperature cycle. Randomly fluctuating temperature regime mortality curves also resembled constant 20°C tanks with mortalities after high temperature exposures (25°C - 30°C). Some test animals survived in all assays with the exception of the 30°C assays, with highest survival associated with low temperature and low Ni concentration. Post-exposure mortality occurred most frequently in individuals which had experienced high Ni concentrations and high temperatures during assays. Additional temperature stress imposed 2 - 12 weeks post exposure resulted in a single death out of 116 individuals suggesting that survivors are capable of surviving subsequent temperature stresses. These investigations suggest that temperature significantly and markedly affects acute nickel toxicity under both constant and fluctuating temperature regimes and plays a role in post exposure mortality and subsequent stress response.

The role of sleep and dreaming in the processing of episodic memory

La présente thèse examine les liens entre le sommeil, la mémoire épisodique et les rêves. Dans une première étude, nous utilisons les technologies de la réalité virtuelle (RV) en liaison avec un paradigme de privation de sommeil paradoxal et de collecte de rêve en vue d'examiner l'hypothèse que le sommeil paradoxal et le rêve sont impliqués dans la consolidation de la mémoire épisodique. Le sommeil paradoxal a été associé au rappel des aspects spatiaux des éléments émotionnels de la tâche RV. De la même façon, l'incorporation de la tâche RV dans les rêves a été associée au rappel des aspects spatiaux de la tâche. De plus, le rappel des aspects factuels et perceptuels de la mémoire épisodique, formé lors de la tâche VR, a été associé au sommeil aux ondes lentes. Une deuxième étude examine l'hypothèse selon laquelle une fonction possible du rêve pourrait être de créer de nouvelles associations entre les éléments de divers souvenirs épisodiques. Un participant a été réveillé 43 fois lors de l'endormissement pour fournir des rapports détaillés de rêves. Les résultats suggèrent qu'un seul rêve peut comporter, dans un même contexte spatiotemporel, divers éléments appartenant aux multiples souvenirs épisodiques. Une troisième étude aborde la question de la cognition lors du sommeil paradoxal, notamment comment les aspects bizarres des rêves, qui sont formés grâce aux nouvelles combinaisons d'éléments de la mémoire épisodique, sont perçus par le rêveur. Les résultats démontrent une dissociation dans les capacités cognitives en sommeil paradoxal caractérisée par un déficit sélectif dans l'appréciation des éléments bizarres des rêves. Les résultats des quatre études suggèrent que le sommeil aux ondes lentes et le sommeil paradoxal sont différemment impliqués dans le traitement de la mémoire épisodique. Le sommeil aux ondes lentes pourrait être implique dans la consolidation de la mémoire épisodique, et le sommeil paradoxal, par l'entremise du rêve, pourrais avoir le rôle d'introduire de la flexibilité dans ce système mnémonique.

Efficacité et toxicité de l'eugénol administré à des doses anesthésiantes chez des grenouilles Xenopus laevis.

L’eugénol permet d’induire une anesthésie chirurgicale chez la grenouille africaine à griffes (Xenopus laevis) sans causer de lésions chez des grosses grenouilles (90-140g). Le premier objectif de la présente étude était de déterminer la durée de l’anesthésie et d’évaluer la dépression du système nerveux central ainsi que les changements de saturation en oxygène et de fréquence cardiaque chez des petites (7.5 ± 2.1 g) et moyennes (29.2 ± 7.4 g) grenouilles Xenopus laevis en fonction du temps d’exposition à un bain d’eugénol de 350 µL/L. Suite à une immersion de 5 ou 10 minutes, la réponse au test à l’acide acétique, au réflexe de retrait et au réflexe de retournement était absente pendant 1 heure (petites grenouilles) et 0,5 heure (moyennes) et l’anesthésie chirurgicale durait au maximum 15 et 30 minutes chez les petites et moyennes grenouilles respectivement. La saturation en oxygène n’était pas affectée de façon significative, mais la fréquence cardiaque était diminuée jusqu’à 1 heure post-immersion dans les deux groupes. Le deuxième objectif était de déterminer la toxicité de l’eugénol chez des grenouilles de taille moyenne après une ou trois administrations à une dose anesthésique, avec ou sans période de récupération d’une semaine. Histologiquement, il y avait de l’apoptose tubulaire rénale et des membranes hyalines pulmonaires après une administration, et de la nécrose hépatique et des hémorragies dans les tissus adipeux après trois administrations. Ces résultats suggèrent que le poids corporel est un paramètre important à considérer lors de l’anesthésie de grenouilles Xenopus laevis par immersion dans l’eugénol.

Evaluation of malacosporean life cycles through transmission studies

Myxozoans, belonging to the recently described Class Malacosporea, parasitise freshwater bryozoans during at least part of their life cycle, but no complete malacosporean life cycle is known to date. One of the 2 described malacosporeans is Tetracapsuloides bryosalmonae, the causative agent of salmonid proliferative kidney disease. The other is Buddenbrockia plumatellae, so far only found in freshwater bryozoans. Our investigations evaluated malacosporean life cycles, focusing on transmission from fish to bryozoan and from bryozoan to bryozoan. We exposed bryozoans to possible infection from: stages of T bryosalmonae in fish kidney and released in fish urine; spores of T bryosalmonae that had developed in bryozoan hosts; and spores and sac stages of B. plumatellae that had developed in bryozoans. Infections were never observed by microscopic examination of post-exposure, cultured bryozoans and none were detected by PCR after culture. Our consistent negative results are compelling: trials incorporated a broad range of parasite stages and potential hosts, and failure of transmission across trials cannot be ascribed to low spore concentrations or immature infective stages. The absence of evidence for bryozoan to bryozoan transmissions for both malacosporeans strongly indicates that such transmission is precluded in malacosporean life cycles. Overall, our results imply that there may be another malacosporean host which remains unidentified, although transmission from fish to bryozoans requires further investigation. However, the highly clonal life history of freshwater bryozoans is likely to allow both long-term persistence and spread of infection within bryozoan populations, precluding the requirement for regular transmission from an alternate host.

In vitro colonic metabolism of coffee and chlorogenic acid results in selective changes in human faecal microbiota growth

Coffee is a relatively rich source of chlorogenic acids (CGA), which, like other polyphenols are postulated to exert preventative effects against cardiovascular disease and type-2 diabetes. As a considerable proportion of ingested CGA reaches the large intestine, CGA may be capable of exerting beneficial effects in the large gut. Here we utilise a stirred, anaerobic, pH controlled, batch culture fermentation model of the distal region of the colon in order to investigate the impact of coffee and CGA on the growth of the human faecal microbiota. Incubation of the coffee with the human faecal microbiota led to the rapid metabolism of CGA (4h) and the production of dihydrocaffeic acid and dihydroferulic acid, whilst caffeine remained un-metabolised. The coffee with the highest levels of CGA (p<0.05, relative to the other coffees) induced a significant increase in Bifidobacterium spp. relative to the control at 10 hours post exposure (p<0.05). Similarly, an equivalent quantity of CGA (80.8mg; matched with that in high CGA coffee) induced a significant increase in Bifidobacterium spp. (p<0.05). CGA alone also induced a significant increase in the Clostridium coccoides-Eubacterium rectale group (p<0.05). This selective metabolism and subsequent amplification of specific bacterial populations could be beneficial to host health.

The neurotoxicity of 5-S-cysteinyldopamine is mediated by the early activation of ERK1/2 followed by the subsequent activation of ASK1/JNK1/2 pro-apoptotic signalling

Parkinson's disease is characterized by the progressive and selective loss of dopaminergic neurons in the substantia nigra. It has been postulated that endogenously formed CysDA (5-S-cysteinyldopamine) and its metabolites may be, in part, responsible for this selective neuronal loss, although the mechanisms by which they contribute to such neurotoxicity are not understood. Exposure of neurons in culture to CysDA caused cell injury, apparent 12-48 h post-exposure. A portion of the neuronal death induced by CysDA was preceded by a rapid uptake and intracellular oxidation of CysDA, leading to an acute and transient activation of ERK2 (extracellular-signal-regulated kinase 2) and caspase 8. The oxidation of CysDA also induced the activation of apoptosis signal-regulating kinase 1 via its de-phosphorylation at Ser967, the phosphorylation of JNK (c-Jun N-terminal kinase) and c-Jun (Ser73) as well as the activation of p38, caspase 3, caspase 8, caspase 7 and caspase 9. Concurrently, the inhibition of complex I by the dihydrobenzothiazine DHBT-1 [7-(2-aminoethyl)-3,4-dihydro-5-hydroxy-2H-1,4-benzothiazine-3-carboxylic acid], formed from the intracellular oxidation of CysDA, induces complex I inhibition and the subsequent release of cytochrome c which further potentiates pro-apoptotic mechanisms. Our data suggest a novel comprehensive mechanism for CysDA that may hold relevance for the selective neuronal loss observed in Parkinson's disease.

Evaluation of cytokines concentration and percentage of survival of rabies virus-infected mice submitted to anti-rabies Vero-cell propagated vaccine and P-acnes

Previously, survival of rabies infection was shown to correlate with low IL-6 serum concentration in mice subjected to post-exposure treatment with the Fuenzalida Palacios rabies vaccine in conjunction with the immunomodulator Propionibacterium acnes, previously Corynebacterium parvum. Considering the substitution of the Fuenzalida Palacios rabies vaccine by the Vero cell raised anti-rabies vaccine in almost all countries, the objective of this work was to evaluate the survival and cytokine serum concentration of rabies virus-infected mice treated with P. acnes in conjunction with or the anti-rabies-VERO vaccine. For this, Swiss mice were experimentally infected with street rabies virus and subjected to vaccine and/or P. acnes following infection. Animals were killed at different times and serum was collected to evaluate cytokines. The greatest survival was observed in animals given one or two does of P. acnes in the absence of vaccination. Animals given anti-rabies VERO vaccine alone or with three doses of P. acnes had the second highest survival rate. The group that had the highest percentage of mortality also had the highest IL-6 concentration on the 10th day, a time correlating with clinical symptoms of the animals. The results reinforce the inefficacy of anti-rabies vaccine in only one dose as a post-exposure treatment irrespective of the type of vaccine used, the immunomodulation activity of P. acnes in rabies post-exposure treatment and suggest a role for IL-6 in rabies virus pathogenesis. (c) 2006 Elsevier B.V. All rights reserved.

Avaliação do efeito da toxina épsilon do clostridium perfringens em monocamadas de células MDCK (Madin-Darby canine kidney cell)

Pós-graduação em Ciência Animal - FMVA

Eficácia pós-expositiva de RNAs de Interferência (siRNAs) e Ribavirina em camundongos infectados com o vírus rábico de origem canina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Proteção imunológica contra raiva em população rural exposta à epidemia em 2005

No mês de maio de 2005, no município de Augusto Correa, nordeste do Estado do Pará, foi diagnosticado um surto de raiva paralítica transmitida por morcegos hematófagos que resultou em 15 casos confirmados da doença. A população exposta recebeu vacinação anti-rábica em regime de pré e pós-exposição, sendo atendidas aproximadamente 3500 pessoas. O presente estudo verificou a persistência de anticorpos neutralizantes antivírus rábico, em amostra da população de Augusto Correa, vacinada com vacina de cultivo celular (Verorab®) por ocasião do surto. Foram coletadas e analisadas 505 amostras de moradores de quatro comunidades de Augusto Correa (Arai, Porto do Campo, Cachoeira e Nova Olinda) em junho de 2007. Os níveis de anticorpos neutralizantes foram dosados através da técnica de Favoretto e relacionados com dados demográficos, com história prévia de malária, novas agressões por animais e com o esquema vacinal recebido. Após dois anos da campanha vacinal, os resultados revelaram a persistência de anticorpos neutralizantes em níveis adequados em 90.5% da população vacinada, em esquemas de pré e pós-exposição, mesmo naqueles que receberam tratamento incompleto. Os níveis de anticorpos neutralizantes não foram reduzidos em função das variáveis analisadas.

Perfil epidemiológico de acidentes com material biológico entre estudantes de medicina em um pronto-socorro cirúrgico

OBJECTIVE: To evaluate the accidents with biological material among medical students interning in a trauma emergency room and identify key related situations, attributed causes and prevention. METHODS: we conducted a study with a quantitative approach. Data were collected through a questionnaire applied via internet, with closed, multiple-choice questions regarding accidents with biological material. The sample comprised 100 students. RESULTS: thirty-two had accidents with biological material. Higher-risk activities were local anesthesia (39.47%), suture (18.42%) and needle recapping (15.79%). The main routes of exposure to biological material were the eyes or mucosa, with 34%, and syringe needle puncture, with 45%. After contamination, only 52% reported the accident to the responsible department. CONCLUSION: The main causes of accidents and routes of exposure found may be attributed to several factors, such as lack of training and failure to use personal protective equipment. Educational and preventive actions are extremely important to reduce the incidence of accidents with biological materials and improve the conduct of post-exposure. It is important to understand the main causes attributed and situations related, so as general and effective measures can be applied.

Atendimento antirrábico humano pós-exposição: proposta de intervenção e estudo da percepção do comportamento de cães e gatos envolvidos nos agravos

Pós-graduação em Medicina Veterinária - FCAV

Entendendo o comportamento canino: estudo das causas de agressão e sua influência na profilaxia da raiva humana

Pós-graduação em Medicina Veterinária - FCAV

Ribavirin has an in vitro antiviral effect in rabies virus infected neuronal cells but fails to provide benefit in experimental rabies in mice

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Relationship Between BP (Baculovirus penaei) Energy Reserves in Larval and Postlarval Pacific White Shrimp Penaeus vannamei

The relationship between energy reserves of the penaeid shrimp Penaeus vannamei and Baculovirus penaei, or BP, were investigated in a series of experiments using mysis stage or early postlarval shrimp. Pre-exposure and post-exposure levels of protein and triacylgycerol (TAG) were determined. The effect of pre-exposure protein and TAG levels on susceptibility to BP infections was also investigated by starving a group of shrimp immediately prior to BP exposure. There was no consistent relationship between either pre-exposure or post-exposure protein levels and the percent of shrimp developing patent BP infections. There was, however, a significant positive correlation between TAG levels immediately prior to viral exposure and prevalence of infection 72 h later. Experimental reduction of TAG reserves prior to BP exposure delayed the development of a patent infection. In some, but not all, experiments there was a significant reduction in TAG levels of infected compared with uninfected shrimp 72 h post-exposure. The effect of patent BP infections on host TAG levels was subordinate to fluctuations in TAG content associated with the ontogeny of the hepatopancreas. Results of this study support histological observations that shrimp lipid levels can be altered by baculovirus infections. Furthermore, high levels of energy reserves in the form of TAG are associated with increased susceptibility to BP infection in larval and postlarval shrimp.