Вы находитесь на странице: 1из 79
Universidade Federal de Santa Maria (UFSM) International Stress and Behavior Society (ISBS) Final Program and
Universidade Federal de Santa Maria (UFSM) International Stress and Behavior Society (ISBS) Final Program and

Universidade Federal de Santa Maria (UFSM) International Stress and Behavior Society (ISBS)

Final Program and Abstracts

UFSM-ISBS JOINT WORKSHOP OF NEUROPSYCHOPHARMACOLOGY, TOXICOLOGICAL BIOCHEMISTRY, AND BEHAVIORAL BIOLOGY

TOXICOLOGICAL BIOCHEMISTRY, AND BEHAVIORAL BIOLOGY Anfiteatro do Colégio Politécnico Universidade Federal de

Anfiteatro do Colégio Politécnico Universidade Federal de Santa Maria December 12-13, 2016 Santa Maria, RS, Brazi

FINAL PROGRAM ORGANIZING COMMITTEE Prof. Denis B. Rosemberg (UFSM, Brazil) President of the Local Organizing

FINAL PROGRAM

ORGANIZING COMMITTEE

Prof. Denis B. Rosemberg (UFSM, Brazil) President of the Local Organizing Committee

Prof. Allan V. Kalueff (St Peterburg State University, Russia, ZNRC Chair) ISBS President

Ana Lúcia A. Segatto, Ph.D. Assistant of Laboratory, UFSM, Brazil

Barbara D. Fontana (UFSM, Brazil) MSc. Student of the Graduate Program in Biological Sciences: Toxicological Biochemistry

Cláudia S. Oliveira, Ph.D. (UFSM, Brazil) PhD Graduate Program in Biological Sciences: Toxicological Biochemistry

Fabiano V. Costa, MSc. (UFSM, Brazil) Graduate Program in Pharmacology

Flavia V. Stefanello (UFSM, Brazil) Scientific Initiation Scholarship Program (CNPq Fellowship), Laboratory of Experimental Neuropsychobiology,

Jamile S. Bernardi (UFSM, Brazil) MSc. Student of the Graduate Program in Biological Sciences: Toxicological Biochemistry

Julia C. Marion (UFSM, Brazil) Scientific Initiation Scholarship Program (FAPERGS Fellowship), Laboratory of Experimental Neuropsychobiology

Luiz Vinícius C. Rosa (UFSM, Brazil) MSc. Student of the Graduate Program in Biological Sciences: Toxicological Biochemistry

Nathana J. Mezzomo, MSc. (UFSM, Brazil) PhD. Student of the Graduate Program in Pharmacology

Paola R. Ziani (UFSM, Brazil) MSc. Student of the Graduate Program in Biological Sciences: Toxicological Biochemistry

Talise E. Müller, MSc. (UFSM, Brazil) PhD. Student of the Graduate Program in Biological Sciences: Toxicological Biochemistry

Vanessa A. Quadros, MSc. (UFSM, Brazil) PhD. Student of the Graduate Program in Biological Sciences: Toxicological Biochemistry

FINAL PROGRAM SPONSORS  International Stress and Behavior Society (ISBS)  Graduation Program in Biological

FINAL PROGRAM

SPONSORS

International Stress and Behavior Society (ISBS)

Graduation Program in Biological Sciences: Toxicological Biochemistry (UFSM)

Graduation Program in Pharmacology (UFSM)

Study English

LabSul

Sorriso’s Gastronomia

MAIN TOPICS

Neuropsychopharmacology and Toxicology

Pharmacological Strategies for Neuroprotection

Experimental Models of Behavior

Neurobiology and Physiology of Stress, Fear, and Anxiety

Pain and Nociception

Learning and Memory

Translational Research in Biological Psychiatry

Biomarkers of Oxidative Stress in the CNS

Focus: The 2-day workshop aimed to exchange and share the developing translational knowledgebase of studies associates to three main topics of interest: Neuropsychopharmacology, Toxicological Biochemistry, and Behavioral Biology, with the particular focus of some mammalian and non- mammalian model organisms in scientific research. Students from undergraduate, graduation, and researchers interested in neurochemistry, biochemistry, pharmacology, physiology, molecular biology and animal behavior were welcome to join the event.

FINAL PROGRAM CONFERENCES Day 1. Monday, December 12, 2016 8:30-9:30 REGISTRATION 9:30-9:45 OPENING CEREMONY

FINAL PROGRAM

CONFERENCES

Day 1. Monday, December 12, 2016

8:30-9:30

REGISTRATION

9:30-9:45

OPENING CEREMONY Paulo A. Burmann (Dean, UFSM) Félix A. A. Soares (Vice Director, CCNE, UFSM) Denis B. Rosemberg (President of the Organizing Commitee, UFSM)

9:45-10:00

LECTURE FROM THE SPONSOR STUDY ENGLISH Ptolomeu Palma

10:00-10:30 COFFEE BREAK

10:30-11:30 MAIN CONFERENCE Allan V. Kalueff (St Peterburg State University, Russia, ISBS President) - Endophenotypes and modeling disease syndromes across taxa - lessons from rodents and zebrafish (tribute to Irv Gottesman)

11:30-12:00 OPEN FOR DISCUSSION

12:00-14:00 LUNCH

14:00-15:15 ROUND

TABLE

CONFERENCE

I:

NON-MAMMALIAN

MODELS

FOR

TOXICOLOGICAL AND BEHAVIORAL NEUROSCIENCE STUDIES

Speaker 1: Eduardo P. Rico (UNESC) - Fish embryo test (FET): Applicability for toxicological research Speaker 2: Denis B. Rosemberg (UFSM) - Neuropsychopharmacology of taurine and alarm reactions in zebrafish Speaker 3: Leticia Arantes (UFSM) - Modeling neurological disorders in Caenorhabditis elegans

15:15-15:30 OPEN FOR DISCUSSION

15:30-16:00 COFFEE BREAK

16:00-17:00 POSTER SESSION I: TOXICOLOGICAL BIOCHEMISTRY

17:00 FREE TIME TO VISIT THE UFSM FACILITIES

FINAL PROGRAM Day 2. Tuesday, December 13, 2016 8:30-9:45 NEUROPSYCHIATRIC DISORDERS ROUND TABLE CONFERENCE II:

FINAL PROGRAM

Day 2. Tuesday, December 13, 2016

8:30-9:45

NEUROPSYCHIATRIC DISORDERS

ROUND

TABLE

CONFERENCE

II:

TRANSLATIONAL

MODELS

OF

Speaker 1: Marilise E. Burger (UFSM) - Neurobiological bases of addiction:

new experimental perspectives Speaker 2: Mauro S. Oliveira (UFSM) - Drug repositioning for the treatment of epilepsy and associated comorbities

9:45-10:00

OPEN FOR DISCUSSION

10:00-10:30 COFFEE BREAK

10:30-11:45 ROUND TABLE CONFERENCE III: NEUROCHEMICAL MECHANISMS OF PAIN AND NOCICEPTION

Speaker 1: Juliano Ferreira (UFSC) - Role of spinal calcium channels in pain Speaker 2: Sara M. Oliveira (UFSM) - Participation of kinins in paclitaxel- induced neuropathy Speaker 3: Gabriela Trevisan (UFSM) - Role of TRP channels in chemotherapy-induced neuropathy

11:45-12:00 OPEN FOR DISCUSSION

12:00-14:00 LUNCH

14:00-15:15 ROUND TABLE CONFERENCE IV: NEUROPROTECTIVE EFFECTS OF SELENIUM-DERIVED COMPOUNDS IN TOXICOLOGICAL BIOCHEMISTRY

Speaker 1: Andreza F. de Bem (UFSC) - Hypercholesterolemia as a risk factor for neurocognitive impairments: Therapeutic approach of organoselenium compounds Speaker 2: Cláudia S. de Oliveira (UFSM) - Mercury: interaction with Selenium Speaker 3: João Batista T. Rocha (UFSM) - Selenium as a neuroprotective agent

15:15-15:30 OPEN FOR DISCUSSION

15:30-16:00 COFFEE BREAK

FINAL PROGRAM 16:00-17:00 POSTER SESSION II : NEUROPSYCHOPHARMACOLOGY AND BEHAVIORAL BIOLOGY 17:00-17:15 FINAL REMARKS

FINAL PROGRAM

16:00-17:00 POSTER SESSION II: NEUROPSYCHOPHARMACOLOGY AND BEHAVIORAL BIOLOGY

17:00-17:15 FINAL REMARKS

19:00 FAREWELL CONFRATERNIZATION (SORRISO'S GASTRONOMIA)

FINAL PROGRAM ABSTRACTS SESSION: TOXICOLOGICAL BIOCHEMISTRY

FINAL PROGRAM

ABSTRACTS

FINAL PROGRAM ABSTRACTS SESSION: TOXICOLOGICAL BIOCHEMISTRY

SESSION: TOXICOLOGICAL BIOCHEMISTRY

PROTECTIVE EFFECTS OF QUERCETIN ON REACTIVE OXYGEN SPECIES IN RATS SUBMITTED TO EXPERIMENTAL ADJUVANT ARTHRITIS

PROTECTIVE EFFECTS OF QUERCETIN ON REACTIVE OXYGEN SPECIES IN RATS SUBMITTED TO EXPERIMENTAL ADJUVANT ARTHRITIS MODEL

Alessandra G. Manzoni 1 , Renata S. Pereira 1,2 , Lívia G. Castilhos 1,2 , Juliana Sorraila 3 , Lara Vargas Becker 1,3 , Karine Lanes Silveira 1,2 , Daniela B. R. Leal 1,2,3

1 Laboratório de Imunobiologia Experimental e Aplicada, Universidade Federal de Santa Maria.

2 Programa de Pós-Graduação em Ciências Farmacêuticas:Análises Clínicas, Universidade Federal de Santa Maria.

3 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: Rheumatoid arthritis (RA) is the most common inflammatory rheumatic disease that affect almost 1% of the world population. The free radicals and oxidative stress are important factors in the development of arthritis. Substances that modify the production of these factors and consequently reduce the extension of tissue damage will be effective in combating inflammation. Some studies have demonstrated an association between RA and oxidative stress. Reactive oxygen species (ROS) have an important role in inflammation induction and tissue damage. Quercetin (QUE) is a flavonoid found in many fruits and vegetables. It has a wide range of biological actions including antioxidant, anti-carcinogenic and anti- inflammatory activities, as well is able to attenuate lipid peroxidation. Objective: The aim of this study was to investigate whether quercetin treatment alters the ROS levels in serum, liver and kidney of rats submitted to a model of induced arthritis. Methods: The animals were divided in ten groups (n=5) and individually treated with QUE (5, 25, 50 mg/kg) or saline, during 45 days and treated with dexamethasone (0.2 mg/kg) during 7 day, as a positive control. The 2′-7′-dichlorofluorescein fluorescence assay was used to measure cellular peroxide production and other reactive species. The Animal Ethics Committee from the UFSM approved all animal procedures (protocol under number: 9970081214). Results: QUE 25 and 50 mg/kg was able to reduce ROS levels in serum, however only the concentration of 50 mg/kg was capable to reduce ROS levels in liver. However, no significantly alterations was found in kidney. In addition, DEX group showed increased ROS formation. Conclusion: These results suggest that QUE prevented the increase on ROS formation in serum and liver while DEX increased this marker. Hence, this study may prove the antioxidant effect of quercetin and that this compound is efficient to prevent the oxidative stress induced by arthritis.

Financial Support: CAPES, FAPERGS.

Key words: arthritis; reactive oxygen species; quercetin.

APOPTOTIC AND DNA DAMAGE: A LINK WITH NEUROINFLAMMATION AND DIFFERENT SEIZURES IN EPILEPTIC HUMAN Aline

APOPTOTIC AND DNA DAMAGE: A LINK WITH NEUROINFLAMMATION AND DIFFERENT SEIZURES IN EPILEPTIC HUMAN

Aline Kegler 1 , Eduardo T. Pascotini 1 , Josi Arend 2 , Michele R. Fighera 1,2

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria. 2 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria.

Introduction: Epilepsy is a common chronic neurological disorder characterized by recurrent seizures. Experimental and clinical findings support the role of inflammatory processes in epilepsy. Furthermore, the oxidative stress is known to be triggered by epileptic seizures, and studies also relate the apoptotic pathway in the epileptic process. However, although some studies using experimental models demonstrated a link between epilepsy and apoptosis, the complex signaling pathways leading to neuronal apoptosis in epilepsy have not been completely elucidated. Objective: We aimed to investigate if exist a difference between seizure type and severity with oxidative, inflammatory, apoptotic, and DNA damage parameters in the epilepsy pathophysiology. Methods: Patients with epilepsy (n=43) and health subjects (control group; n=41) were recruited to participate in the study. Initially, the participants have submitted clinic questionnaire and epileptic patients were classified according to their type seizure. Inflammatory and apoptotic factors, oxidative stress parameters, DNA damage, and enzymatic antioxidants were also measured in specific blood samples. We compared these results with data from control subjects (n=41). Results: Statistical analyses showed that patients with epilepsy presented higher levels of TBARS (p<0.05), TNF-α (p<0.05), Caspase 8 (p<0.05), Caspase 3 (p<0.05) and Picogreen (p<0.05) when compared to control subjects. In addition, generalized epilepsy presented higher SOD2 activity when compared to partial seizure and control group. Furthermore, patients with generalized epilepsy had a significant correlation among TNF-α with Caspase 8 (p<0.05), Caspase3 (p<0.05), and Picogreen (p<0.05). We did not observe any significant correlation between the analyzed markers with antiepileptic drugs (AEDs) (p>0.05). Conclusion: Our data suggest that an increase of seizure activity causes changes in the brain, and in relation to generalized crises, these changes are more widespread throughout the nervous system resulting in increased levels of oxidative, inflammatory and apoptotic markers in the peripheral blood.

Financial Support: CNPq, CAPES,

Key words: apoptosis; epilepsy; inflammation.

CENTRAL AND PERIPHERAL NEUROTOXICITY INDUCED BY Rhinella icterica (SPIX, 1824) TOAD VENOM IN Nauphoeta cinerea

CENTRAL AND PERIPHERAL NEUROTOXICITY INDUCED BY Rhinella icterica (SPIX, 1824) TOAD VENOM IN Nauphoeta cinerea COCKROACHES

Allan P. Leal 1 , Raquel O. Soares 1 , Ana Paula A. Perin 1 , Bruna T. Borges 1 , Barbara A.

B.

Ogata 1 , Etiely Karnopp 1 , Maria E. Rosa 1 , Yuri C. Barreto 1 , Ana P. Zanatta 1 , Tiago

G.

dos Santos 3 , Chiara Valsecchi 1 , Lúcia Vinadé 1 , Cháriston André Dal Belo 1,2

1 Laboratório de Neurobiologia e Toxinologia, (LANETOX), Universidade Federal do Pampa. 2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria. 3 Laboratório de Estudos em Biodiversidade Pampiana (LEBIP), Universidade Federal do Pampa.

Introduction: Rhinella icterica is a common venomous toad in Brazil in which the biological activity of its poison has never been described before. Objective: This work investigated the mechanisms of the entomotoxic activity of the methanolic extract of R. icterica venom (MERIV). Methods: Adult Nauphoeta cinerea cockroaches were used as experimental model. The neuromuscular activity of MERIV was evaluated using the in vivo cockroach neuromuscular preparation (CNP), as described elsewhere (Martinelli et al. 2014). Cardiotoxicity was evaluated using the ex situ cockroach semi-isolated heart preparation (CSIHP), by counting the heart rate visually in an eyepiece of a microscope. Grooming behavior was measured in as described in (Carrazoni et al. 2016). The in vivo locomotory activity was filmed in an open field during 10min with a webcam. The movies were then analyzed by the IDtracker® and Matlab® software. All data were expressed as mean ± standard error (SE) and analyzed by Student “t” test or ANOVA followed by Tukey or Dunnett tests. Results: The effect of MERIV (20, 50, and 100µg/g animal weight) on CNP resulted in a significant neuromuscular blockade, in 120min recordings (n=6, p<0.001). The effect of MERIV (1, 2, 4 and 8µg/200µl) on CSIHP was dose-dependent with a maximum decrease of heart rates (50±4 beats/min) for the highest concentration, in 30min (n=9, p<0.05). MERIV (2, 10 and 20 μg/g animal weight) significantly increased grooming activity that was prevented by the previous treatment with phentolamine (0.1μg/g) (n=30, p<0.05 respectively). The same doses significantly decreased the cockroaches locomotory activity, increasing the number of immobility episodes (n=33, p < 0.05). Conclusion: The biological activity of MERIV in cockroaches affects both central and peripheral nervous systems and probably involves the modulation of the octopaminergic transmission.

Financial Support: PRONEM/FAPERGS/CNPq 003/2011 and Edital Toxinologia CAPES 063/2010.

Key words: Toad venom; entomotoxic activity; behavior.

EFFECTS OF METHYLPREDNISOLONE ON RODENT CENTRAL NERVOUS SYSTEM: MODULATION OF CALCIUM-DEPENDENT MEMORY PATHWAYS Barbara A.

EFFECTS OF METHYLPREDNISOLONE ON RODENT CENTRAL NERVOUS SYSTEM: MODULATION OF CALCIUM-DEPENDENT MEMORY PATHWAYS

Barbara A. B. Ogata 1 , Raquel S. Oliveira 1 , Liane Vargas 2 , Allan Pinto Leal¹ , Bruna Trindade Borges¹, Maria Eduarda Rosa¹, Yuri Correia Barreto¹, Ana Paula Zanatta¹, Etiely Karnop¹, Pamela M. Carpes 2 , Cháriston Dal Belo 1 , Lúcia Vinadé 1 .

1 Laboratório de Neurobiologia e Toxinologia, Universidade Federal do Pampa, São Gabriel, RS.

2 Grupo de Pesquisa em Fisiologia, Universidade Federal do Pampa, Uruguaiana, RS.

Introduction: It has been recognized that glucocorticoids have the ability to influence brain cognition. Studies have shown that the glucocorticoid Methylprednisolone (METP) induces a positive modulation of mammalian neuromuscular transmission, which results in increased calcium influx in end plates. We suggest that similar mechanisms are related to calcium signaling involved with the mnemonic response of METP. Objective: The aim this study was to investigate the effects of METP on cell viability and on pathways involved in memory, such as the calcium and calmodulin-dependent kinase type II (CaMKII) in the central nervous system of rodents. Methods: Cell viability of mice cortex and hippocampal brain slices, incubated with METP at 3, 30 and 300 µM, was assayed with the MTT colorimetric test, according to Dal Belo (2013). Viability was also assayed following the model of glucose and oxygen deprivation (GOD) according to Strasser and Fisher (1995). As for in vivo assays, rats were treated with 3 and 300 µM METP for 10 days and its cortex and hippocampal brain structures were homogenized with a TissueLyser (QIAGEN). Pellet and supernatants fractions were submitted to a 10% SDS-PAGE, dyed with Coomassie blue and protein bands (approximately 50kDa) were analyzed by ImageJ (NIH software). All data were expressed as mean ± S.E.M. and significance by Student “t” test. The use of animals was previously approved by the Ethics Commission on Animal Use (CEUA/UNIPAMPA 037/2012). Results: The treatment with 30 µM METP increased the number of viable cells (+45%, p<0.05) on hippocampal slices, while at the lowest dose no difference were found. The cell viability significantly decreased on hippocampal and cortical slices treated with the highest dose (-88% and - 83%, respectively, p<0.05). The results obtained with GOD were not significant, thus the observed increase in cell viability is not related to neuroprotection. The in vivo treatment showed an increase in the density of the 50 kDa electrophoretic band at the lowest dose of METP treatment in both structures (3-4%, p<0.05). Conclusion: These results suggest an enhancement of the CaMKII content and/or in its electrophoretic mobility under treatment with METP at low doses, thus indicating the involvement of a calcium-dependent signaling pathway common in memory-related events.

Financial Support: UNIPAMPA, PRONEM, FAPERGS, CNPq 003/201.

Key words: Glucocorticoids; memory; CaMKII.

EFFECT OF Manilkara rufula EXTRACT ON NEUROMUSCULAR JUNCTION OF COCKROACHES Nauphoeta cinerea Bruna T. Borges

EFFECT OF Manilkara rufula EXTRACT ON NEUROMUSCULAR JUNCTION OF COCKROACHES Nauphoeta cinerea

Bruna T. Borges 1 , Allan P. Leal 1 , Ana P. Zanatta 1 , Barbara A. B. Ogata 1 , Etiely Karnopp 1 , Maria E. Rosa 1 , Yuri C. Barreto 1 , Lúcia Vinadé 1 , Cháriston André Dal Belo 1,2 , Patrícia B. Vieira 1

1 Laboratório de Neurobiologia e Toxinologia (LANETOX), Universidade Federal do Pampa. 2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: Plant secondary metabolites can be applied in several areas of biotechnology, including the development of novel insecticides. Vegetal species adapted to extreme environmental conditions, such as low humidity and high temperatures, present a large diversity of secondary metabolites, which may contribute to their survival. The Caatinga ecosystem in Northeast Brazil is an example of an extreme environment in which most of the native plants have never been subjected to a scientific chemical and biological survey. Objective:

The aim of this work was to evaluate the effect of crude extracts and enriched fractions of Manilkara rufula, a plant from the Caatinga, using the nervous system of cockroaches as a biological model. Methods: Adult Nauphoeta cinerea (Olivier) cockroaches (3-4 months after adult molt) of both sex, were used. The neuromuscular activity was evaluated using the in vivo cockroach neuromuscular preparation (CNP), as described elsewhere (Martinelli et al. 2014, Biochimica et Biophysica Acta 1840, 935-944). The treatments were made injecting in the third abdominal segment of the animals, 10 µl of the test compounds previously dissolved in physiological solution. The crude extract of M. rufula (CEMR) was administered at 50, 100 and 200 µg per animal and the methanolic (MF) and aqueous (AF) fractions were administrated at 10, 20 and 40 µg per animal. Results: Overall the application of the CEMR induced a time-dependent decrease of the muscle twitch tension that was the maxim for 100 µg/g of animal weight, in 120 min. When the MF and AF were assayed there was a similar effect to the crude extract that was the maxim for the highest concentration. Discussion and conclusion: The results confirm the entomotoxic potential of Manilkara rufula extract. Phytochemical and guided biological assays are in the course and will be able to identify the chemical compounds related to the toxic effects in insects.

Key words: Natural insecticides; Nauphoeta cinerea; neuromuscular junctions.

CHRONIC EXPOSURE TO METHYLMERCURY CAUSES ENZYMATIC ALTERATIONS IN THE COCKOROACH Nauphoeta cinerea Bruna Candia Piccoli

CHRONIC EXPOSURE TO METHYLMERCURY CAUSES ENZYMATIC ALTERATIONS IN THE COCKOROACH Nauphoeta cinerea

Bruna Candia Piccoli 1 , Cláudia Sirlene de Oliveira 1 , Fernanda D’Ávila da Silva 1 , Jéssica da Costa Alvim 2 , João Batista Teixeira da Rocha 1,2

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

2 Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Santa Maria.

Introduction: Mercury is a toxic metal, widespread in the environment. There are different forms of mercury in the environment, for instance, elemental mercury (Hg 0 ), inorganic (Hg 2+ ), and organic (mainly MeHg). MeHg is synthetized by the aquatic microorganisms through methylation of inorganic mercury. In this way, MeHg is bioaccumulated in the aquatic food chain, with the piscivorous fish reaching great amount of this metal. Once ingested by the organisms, MeHg is easily absorbed in the gut and is transported to tissues mainly to the central nervous system. The high toxicity of MeHg is due to the fact that it is a soft electrophile and in the living organism have great affinity for soft nucleophile, for example, thiol (-SH) and selenol (-SeH) groups ubiquitously distributed in the cells. Objective: The aim of this study was to investigate whether chronic exposition to MeHg alters antioxidant system and acetylcholinesterase activity. Methods: The experiments were divided in two protocols: 1) Cockroaches were fed with different concentrations of MeHg (0, 2.5, 5, 25 and 100 µg/g of food) during three months; 2) Cockroaches were fed with different concentrations of MeHg (0, 2.5, 5, 25 and 100 µg/g) during thirty days and, after this period, started to receive diet without MeHg during two months. After the treatments, cockroaches were euthanized and the head and fatty body were collected. Production of reactive oxygen species, glutathione S-transferase and acetylcholinesterase activity were evaluated. Results Treatment with MeHg did not alter the reactive oxygen species production in the tissues evaluated in both protocols. Only the cockroaches from protocol 1 presented biochemical alterations. For instance, the exposure to the highest concentration of MeHg inhibited acetylcholinesterase activity in head and fatty body. Moreover, the highest concentration of MeHg caused an increased in glutathione S-transferase activity in head and the two highest concentrations of MeHg increased this enzyme activity in the fatty body. Conclusion: Cockroaches are a good model to study the exposition to MeHg. Our data suggested that even in low concentrations MeHg alters the activity of important enzymes.

Financial Support: CAPES, CNPq, FAPERGS.

Key words: cockroaches; behavior; organophosphate; oxime.

MANCOZEB EXPOSURE DURING LARVAL AND ADULT PHASE AFFECTS ANTIOXIDANT ENZYMES ACTIVITY AND IMPAIRS LOCOMOTOR PERFORMANCE

MANCOZEB EXPOSURE DURING LARVAL AND ADULT PHASE AFFECTS ANTIOXIDANT ENZYMES ACTIVITY AND IMPAIRS LOCOMOTOR PERFORMANCE OF Drosophila melanogaster

Cynthia Camila Ziech 1 , Eduardo da Rosa Ávila 1 , Nathane Rosa Rodrigues 2 , Giulianna Echeverria Macedo 1 , Gustavo Felipe da Silva 1 , Karen Kich Gomes 1 , Jessica Ferreira Rodrigues¹, Jeferson Luis Franco 1 , Thaís Posser 1

1 Oxidative Stress and Cell Signaling Research Group (GPEOSCEL), Universidade Federal do Pampa, Campus São Gabriel, 97300-000 São Gabriel, RS, Brazil; 2 Departamento de Química, Programa de Pós Graduação em Bioquímica Toxicológica, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brasil;

Introduction: Mancozeb (Manganese ethylene-bis-dithiocarbamate polymeric complex with zinc salt) is an antifungal agent largely used in brazilian agriculture mainly in fruits cultivation. Environmental and occupational exposure of humans to this agent is related with symptoms associated with a neurological pathology called Manganism, presenting clinical and biochemical markers very similar to Parkinson’s Disease. Differential susceptibility to exposure to pesticides is observed in developmental and adult phase in mammals. However, little is known about Mancozeb toxicity in different developmental stages. Drosophila melanogaster is an alternative model for toxicological studies that has a short period of life, this character is very useful for studies of effects of chemical compounds on different stages of development. Objectives: In this study, we investigate the effects of Mancozeb consumption in adult and larval phase of D. melanogaster, on antioxidant enzymes activity and locomotor ability. Methods: For larval exposure protocol, eggs were ovoposited and larves ecloded and grown in medium containing 0, 0.1 and 1 mg/mL of Mancozeb. For adult exposure protocol, 1-3 days old flies were exposed up to 15 days to medium containing Mancozeb (1-10 mg/mL). Locomotor behavior was analyzed by negative geotaxis. For glutathione-S-transferase (GST) and catalase activity (CAT), 20 flies were homogeneized in Tris 20mM Ph7.4 and enzymes activity measured in supernatant. Results: The ingestion of Mancozeb 1 mg/mL during larval developmental phase, impaired locomotor performance of newly ecloded flies, delaying the time to reach 5 cm vertically in a glass tube (from 3.15 ± 1.01 s to 7.69 ± 1.9 s). GST activity was inhibited in 42% in relation to control. No alteration on CAT activity was observed. Adult flies were more resistant to exposure to Mancozeb, locomotor deficit was observed only after 15 days of exposure to 1 mg/mL. Catalase activity was inducted in 169% from 10 mg/mL; GST activity was induced in 106% after 5 mg/mL. No effects were observed in lower concentrations. Conclusion: the toxicity of Mancozeb was more prominent in the protocol of exposure during larval phase of D. melanogster. Oxidative stress and damage to Central and Peripheral Nervous System are targets potentially affected by this compound in larval and adult phase.

Financial Support: CNPq, CAPES, FAPERGS, Unipampa

Key words: Drosophila melanogaster; Mancozeb; antioxidant enzymes.

CAFFEINE PROMOTES ADAPTIVE RESPONSE OF BRAIN BIOENERGETICS AFTER THE NEUROTOXICITY INDUCED BY ACEAMINOPHEN INTOXICATION

CAFFEINE PROMOTES ADAPTIVE RESPONSE OF BRAIN BIOENERGETICS AFTER THE NEUROTOXICITY INDUCED BY ACEAMINOPHEN INTOXICATION

Débora Gonçalves 1 , Nelson R. de Carvalho 2 , Aline Courtes 1 , Pamela Carvalho 1 , Ingrid Kich 1 , Félix A. Soares 1 , Thais Poser 2 , Jeferson L. Franco 2 .

1 Departamento de Bioquímica e Biologia Molecular, Centro de Ciências Naturais e Exatas, Universidade Federal de Santa Maria, Campus UFSM, Santa Maria, RS, Brazil. 2 Campus São Gabriel, Universidade Federal do Pampa, São Gabriel, RS, Brazil.

Introduction: Neurotoxicity associated with acetaminophen (APAP) intoxication is closely related to the mitochondrial dysfunction. So, natural compounds, such as caffeine, could have a protective effect in the mitochondrial oxidative phosphorylation (OXPHOS) in the brain, which remains unclear. In this context, the OXPHOS could have been extensively explored by High-Resolution Respirometry (HRR) demonstrating to be a useful methodology to study the bioenergetics parameters, helping to understand the mechanisms in the survival and maintenance of normal biological function. Thus, in order to understand the mechanism of caffeine and APAP during the neurotoxicity, this work aims to evaluate the changes in the mitochondrial bioenergetics function in the brain through the high-resolution respirometry during the neurotoxicity associated with APAP intoxication. Methods: Seven-week-old male adult Swiss albino mice (30 40g) were randomly divided into four groups: Control (saline 0.9%; 20 mL/Kg), Caffeine (Caf; 20 mg/Kg), APAP-treated (250 mg/Kg; 15 mL/Kg) and Caf+APAP. All the solutions were administered by the intraperitoneal (i.p.). Animals were sacrificed after 4 h. Brain homogenate (0.1 mg/mL) was used to the HRR in Oxygraph-2k (O2k, OROBOROS Instruments, Innsbruck, Austria). All experiments were performed at 37 o C using DatLab 4.0 software (Oroboros Inc., Austria), with continuous stirring at 750 rpm. HRR analyses coupled to substrate- uncoupler-inhibitor titration (SUIT) protocols established in the literature. This study was approved by the Ethics and Animal Welfare Committee of Federal University of Santa Maria, Brazil (Permit Number:

3208150915). Significance was assessed by two-way analysis of variance (ANOVA), followed by Newman– Keuls’s Test for post-hoc comparison. Values of p < 0.05 were considered statistically significant. Results:

Basal respiration and Complex I (glutamate and malate) leak remained unchanged in all groups. HRR assay demonstrated that the Caffeine and Caf+APAP groups were able to increase of OXPHOS when the convergence pathway was stimulated by substrates of Complex I and Complex II (succinate). All groups presented a significant increase of OXPHOS stimulated by complex II (succinate and rotenone) when compared to control. In addition, APAP and Caf+APAP groups demonstrated a significant increase in the Rox respiration in relation to control, and only the Caf+APAP group was able to increase the Complex IV respiration. Conclusion: Although the neurotoxicity associated with APAP intoxication is an important factor that contributes to high mortality of APAP overdose, the caffeine and the caffeine associated to APAP promoted an adaptive response in brain mitochondrial bioenergetics parameters analyzed by HRR.

Financial Support: CNPq, CAPES.

Key words: Acetaminophen; Caffeine; High-Resolution Respirometry; Mitochondrial Bioenergetics.

THE BIOLOGICAL ACTIVITY OF Bothriurus bonariensis SCORPION VENOM IN INSECTS AND MAMMALS Etiely Karnopp 1

THE BIOLOGICAL ACTIVITY OF Bothriurus bonariensis SCORPION VENOM IN INSECTS AND MAMMALS

Etiely Karnopp 1 , Douglas S. Santos 1,3 , Raquel S. Oliveira 1 , Allan P. Leal 1 , Ana P. Zanatta 1 , Barbara A. B. Ogata 1 , Bruna T. Borges 1 , Maria E. Rosa 1 , Lucas F. Maciel 1 , Yuri C. Barreto 1 , Lúcia Vinadé 1 , Jaderson C. Costa 2 , Paulo M. Pinto 4 Cháriston A. Dal

Belo 1,2,3

1 Laboratório de Neurobiologia e Toxinologia, (LANETOX), Universidade Federal do Pampa. 2 Instituto do Cérebro (INSCER), Pontifícia Universidade Católica do Rio Grande do Sul. 3 Programa de Pos-Graduação em Ciências Biológicas: Bioquímica Toxicológica. 4 Laboratório de Proteômica Aplicada, Universidade Federal do Pampa.

Introduction: Bothriurus bonariensis is the scorpion most involved in stings in Southern Brazil. Despite of its public health importance, no previous work has been designed to study the B. bonariensis venom (BBV) biological activity. Objective: The aim of this work was to investigate the BBV neurobiological activity using insect and mammalian nervous system preparations. Methods: Central neurotoxicity was evaluated using primary hippocampal cultures prepared from newborns rats, by means of conventional methodology. The cells were used between 10-14 day for Ca 2+ imaging protocols. Peripheral neurotoxicity was accessed by using both the in vivo and the ex situ cockroach metathoracic-coxal adductor muscle preparation and the extracellular leg recordings of spontaneous neural compound action potentials (SNCAP), respectively, that were mounted as described elsewhere (Carrazoni et al. 2016, Toxicology 368, 162-171). Data were expressed as means ± standard deviation (SD) and analyzed by Student “t” test or ANOVA followed by Tukey test. The work was approved by the Ethical Committee in Animal Use (CEUA) UNIPAMPA, under number 043/2015. Results:

BBV (32 µg/g) induced neuromuscular blockade in the in vivo cockroach nerve-muscle preparations (70 ± 4%, n = 6, p < 0.001), causing repetitive twitches and decreasing the frequency of SNCAPs from 82 ± 3.min -1 to 36 ± 1.3.min -1 (n = 6, p< 0.05), without affecting the amplitude. BBV induced a significant increase of Ca 2+ influx (250 ± 1% peak increase, n = 3, p < 0.0001), in primary hippocampal cultures of rats. The co-treatment of hippocampal cells with tetrodotoxin prevented the BBV calcium influx. Conclusion: Our results show that BBV activity is mostly related to a modulation of sodium channels function. This biological activity survey suggests that BBV may have a promising insecticidal and therapeutic potential.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: Scorpion venom; neuromuscular blockade; sodium channels.

PROTECTIVE EFFECTS OF SUPEROXIDE DISMUTASE MIMETIC COMPOUNDS ON OXIDATIVE STRESS IN Caenorhabdiris elegans Fabiane Bicca

PROTECTIVE EFFECTS OF SUPEROXIDE DISMUTASE MIMETIC COMPOUNDS ON OXIDATIVE STRESS IN Caenorhabdiris elegans

Fabiane Bicca Obetine Baptista, Leticia Priscilla Arantes, Alisson Rodrigues Vinholes, Aline Frazen da Silva, Danela de Freitas Camara, Marina Lopes Machado, Tássia Limana da Silveira, Thayanara Cruz da Silva, Félix Alexandre Antunes Soares.

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: Reactive oxygen species (ROS) production is a physiological process necessary for normal cell function. However, in excessive amounts, it can lead to oxidative stress, which is involved in the etiopathogenesis of various diseases and also aging. Superoxide radical are highly reactive oxygen species which can be neutralized to hydrogen peroxide through catalytic action of superoxide dismutase (SOD). The importance of SOD can be demonstrated by the fact that it is the most abundant enzyme of the organism, and it is also the fifth most abundant protein in the same body. Therefore, three SOD mimetic compounds with structural differences were synthesized, in order to be used to prevent oxidative stress-related diseases. The nematode Caenorhabditis elegans is structurally simple but its genome and its metabolic and biosynthetic pathways are highly conserved in mammals. Thus, this model was used in our study. Objective: Investigate the antioxidant activity of SOD mimetic compounds in vivo using C. elegans. Methods: Young adult wild type (N2) C. elegans were exposed to the three SOD mimetics dissolved in DSMO 0.5% (final concentration) at various concentrations for 1 hour in M9 buffer. After washing, the nematodes were exposed to juglone (5- hydroxy-1,4-naphthoquinone) 200 μM, an in vivo superoxide radical generator, for 1 hour, and then scored as dead or alive using a microscope. Results: All SOD mimetic compounds tested significantly increased C. elegans survival to juglone in approximately 30% compared to control. 1and 2showed antioxidant activity starting at 0.50 μM and 3 at 2μM. Conclusion: The SOD mimetic compounds tested showed antioxidant activity in low concentrations and deserve more studies including their effectiveness in oxidative stress-related diseases models.

Financial Support: CAPES, CNPq

Key words: Caenorhabditis elegans; oxidative stress; superoxide dismutase.

SAZONAL VARIATION OF TOXICITY BIOMARKERS IN Loricariichthys anus (Valenciennes, 1835) EXPOSED TO AGRICULTURAL POLLUTION

SAZONAL VARIATION OF TOXICITY BIOMARKERS IN Loricariichthys anus (Valenciennes, 1835) EXPOSED TO AGRICULTURAL POLLUTION

Gustavo H. Weimer 2 , Aline M. B. do Amaral 1,2 , Aline da S. Faria 2 , Carolina L. Gressler 2 , Dionatan de Pellegrin 2 , Gracielle da S. Félix 2 , Letícia K. de Moura 2 , Kamila E. Menezes 2 , Rafaela R. M. de Souza 2 , Vania Lucia Loro 1,2

1 Programa de Pós-Graduação em Biodiversidade Animal, Universidade Federal de Santa Maria 2 Laboratório de Toxicologia Aquática, Universidade Federal de Santa Maria

Introduction: The cropping system based on intensive use of land and constant application of agricultural chemicals inflict a threat to aquatic organisms since these substances will be inevitably carried out to water bodies. The interaction of these agrochemicals with aquatic animals such as fish can lead to a cascade of oxidative reactions with possible damage to cellular components. Moreover, abiotic factors such as precipitation, temperature, and dissolved oxygen can per se cause changes in oxidative status or can act synergistically or antagonistically with agrochemicals. Objective: This study investigated the influence of the intrinsic characteristics of summer and winter in the interaction with agrochemicals on biochemical responses of L. anus, benthic and omnivorous species widely distributed in southern Brazil. Methods: Ten armored catfishes were collected in six points up to downstream in February and August of 2016 in the Passo Real reservoir, a region of intense agricultural production. Agrochemicals were identified and quantified in water and sediment samples. Liver, gills, brain and muscle tissues were analyzed for toxicity parameters. Results:

No pesticides were detected in the sediment. In the water, there were Atrazine, Imidacloprid, Simazine, Azoxystrobin and Propoxur in the winter and only Atrazine in the summer. The most significant difference between seasons was evident in the GST activity in the liver, which increased almost three times in winter at all points. There was no difference between TBARS and carbonyl in liver between seasons. There was a tendency to increase the level of TBARS in brain and gills at some points in the summer, unlike muscle whose values were higher in the winter. AChE activity in brain and muscle increased in winter in nearly all points. Conclusion: As detoxification organ, the liver appears to be more interesting to assess possible effects of agrochemicals on L. anus metabolism. The other organs seem to have been influenced by environmental variables and, therefore, the antioxidant responses could not explain the damage. The enzyme GST appears to have had a protective effect on the liver during the winter where many pesticides have been detected, thus avoiding damage to proteins and lipids.

Financial Support: CAPES, CNPq.

Key words: armored catfish; environmental assessment; oxidative stress.

Manilkara rufula PROTECTS AGAINST LOCOMOTOR PERFORMANCE DAMAGE AND MORTALITY INDUCED BY CHLORPYRIFOS IN Drosophila

Manilkara rufula PROTECTS AGAINST LOCOMOTOR PERFORMANCE DAMAGE AND MORTALITY INDUCED BY CHLORPYRIFOS IN Drosophila melanogaster

Jéssica Ferreira Rodrigues 1 , Karen Kich Gomes 1 , Nathane Rosa Rodrigues 2 , Giulianna Echeverria Macedo 1 , Illana Kemerich Martins 1 , Cynthia Camila Ziech 1 , 1 Thaís Posser, Patrícia de Brum Vieira 1 , Jeferson Luis Franco 1 .

1 Oxidative Stress and Cell Signaling Research Group (GPOSCEL), Universidade Federal do Pampa, Campus São Gabriel, 97300-000 São Gabriel, RS, Brazil; 2 Departamento de Química, Programa de Pós Graduação em Bioquímica Toxicológica, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brasil;

Introduction: Chlorpyrifos (Cp) is an organophosphate insecticide widely used for controlling agricultural and household pests. The main damage caused by Cp is the neurotoxicity mostly attributed to the inhibition of acetylcholinesterase, increasing neurotransmitter acetylcholine in the synaptic cleft and promoting hyperexcitation in central nervous system and muscular junctions. Occupational exposure is the main form of human poisoning by organophosphates and current therapies for these compounds intoxication are not efficient. Therefore the search for compounds acting on organophosphate damage has been intensified. Plants are a potential source of compounds with protective action, as evidenced in literature, acting on several neuropathological conditions. Manilkara rufula is a plant native from Caatinga and Cerrado Biome. Although the pharmacological potential of the genus has been reported, knowledge about this species remains scarce. The biological activities reported for Manilkara species are attributed to secondary metabolites such as triterpene saponins. Drosophila melanogaster is an alternative animal model widely used in toxicological studies. About 75% of disease-related genes in humans have functional orthologs in D. melanogaster and their fast reproductive cycle and easy maintenance make them ideal for use in in vivo bioassays. Objective: This study aims to evaluate the hydroalcoholic extract of leaves of Manilkara rufula (HEMr) antioxidant and protective potential on mortality and locomotor damage induced by Cp in Drosophila melanogaster. Methods:

For treatments, adult female flies (1-4 days old) were exposed to sucrose 1% (control) or HEMr 20 and 50 mg/ml alone or in combination with Cp 0.75 ppm for 24h both diluted in sucrose 1%. Antioxidant activity was measured by analysis of ABTS radical chelation by the extract. Total phenols were measured by Folin- Ciocalteau agent. Finished the treatment, mortality was recorded and surviving flies were submitted to negative geotaxis locomotor test. Results: The HEMr showed ABTS radical sequestration potential (122,9 ± 9,6 µM equivalent to ascorbic acid / 100 mg extract) and satisfactory total phenols content (22.5 ± 2.7 g equivalent to gallic acid / 100 g extract). Exposure of flies to Cp decreased in 44% the surviving of flies and causes impaired locomotor performance in 42%. Both effects were avoided by the extract. Conclusion: The results indicate that M. rufula presents antioxidant potential and protective effect against the toxicity caused by chlorpyrifos on survival and locomotor performance of flies. Further studies are needed to understand the HEMr mechanisms of protection.

Financial Support: CNPq, CAPES, FAPERGS and Unipampa

Key words: Organophosphate toxicity; vegetal extract; antioxidant potential.

Croton campestris METHANOLIC FRACTION PROTECT AGAINST PARAQUAT AND CHLORPYRIFOS INDUCED TOXICITY IN Drosophila

Croton campestris METHANOLIC FRACTION PROTECT AGAINST PARAQUAT AND CHLORPYRIFOS INDUCED TOXICITY IN Drosophila melanogaster

Karen Kich Gomes 1 , Giulianna Echeverria Macedo 1 , Nathane Rosa Rodrigues 1 , Jéssica Ferreira Rodrigues 1 , Cynthia Camila Ziech 1 , Jeferson Luis Franco 1 , Thaís Posser 1 .

1 Grupo de Pesquisa Estresse Oxidativo e Sinalização Celular: Universidade Federal do pampa; Campus São Gabriel, Rio Grande do Sul, Brasil, CEP 97300 000.

Introduction: The exacerbated use of agrochemicals in agriculture is associated with damage to the environment and human health. Among these agrochemicals is Paraquat an herbicide with large spectrum used in a variety of cultures. It was reported Parkinsonism-like syndrome in response to Paraquat exposure and loss of dopaminergic neurons. Chlorpyrifos is an organophosphate insecticide whose toxicity is associated with neurological effects, persistent developmental disorders and autoimmune disorders in humans. In this context, plants can represent a potential source of molecules with pharmacological properties acting on prevention or treatment of illness associated from agrochemical exposure. Croton campestris (pop. velame-do-campo) is a species found in the Brazilian Cerrado and used as a medicinal plant by local population in several morbidities drawing attention for biological properties of this specie. Objective: We aimed to obtain the phytochemical profile of phenolic compound and flavonoids for methanolic fraction of leaves of Croton campestris (MFCC) by HPLC-DAD and to investigate the biologic effect of MFCC against toxicity of Paraquat (PQ) and Chlorpyrifos (CP) in Drosophila melanogaster taking into account viability and locomotor performance. Methods: Female flies (1-4 days) were exposed to MFCC (0, 0.1, 1, 10, 50 mg/mL) mixed to the medium for 7 days for analysis of toxicity of plant. Flies were co-exposed to the medium containing MFCC 1 mg/mL with or without 5mM PQ or 0.25 ppm CP for 48 hours. After the treatment, we evaluate survival rate and locomotor activity by negative geotaxis assay. Results: The chromatography profile of MFCC revealed the presence of organic acids, flobabenc tannis, cumarines, flavones, flavonols, quinoses, flavononols, flavonone and terpenes. MFCC exposure did not present toxicity per se. PQ and CP caused a significant reduction in survival (58% and 34%) respectively. An 80% and 58% decreasing in locomotor activity was observed by PQ and CP respectively. The mortality induced by agrochemicals was completely in the presence of plant. The locomotor performance impairments induced by PQ and CP was partially avoided by plant. Conclusion: Our data shows the protective potential of methanolic fraction of Croton campestris against largely used agrochemicals, whose toxicity is mostly associated with neurotoxic events drawing attention for the biological potential of this species as a source of bioactive components.

Financial Support: CNPq, CAPES, FAPERGS e Unipampa.

Key words: Agrochemicals; Neurodegenerative diseases; Croton campestris.

BIOCHEMICAL PARAMETERS IN GUPPY ( Poecilia vivipara ) EXPOSED TO ZINC IN DIFFERENT ENVIRONMENTAL CONDITIOS

BIOCHEMICAL PARAMETERS IN GUPPY (Poecilia vivipara) EXPOSED TO ZINC IN DIFFERENT ENVIRONMENTAL CONDITIOS

Letícia Kuhn de Moura 1 , Jossiele Wesz Leitemperger 2 , Ana Carolina Gressler 1 , Talise Ellwanger Müller 2 , Bibiana Silveira Moraes 3 Vania Lucia Loro 2,3

1 Graduanda em Zootecnia, Universidade Federal de Santa Maria. 2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria. 3 Programa de Pós-Graduação em Ciências Biológicas: Biodiversidade Animal, Universidade Federal de Santa Maria.

Introduction: Contamination of aquatic ecosystems by metals causes various biochemical changes in aquatic organisms, and fish are recognized as indicators of environmental quality. Zinc (Zn) is a transition metal which participates in the modulation of regulatory proteins and cellular activities. Poecilia vivipara (guppy) is a Brazilian euryhaline teleost fish is commonly found in both fresh and coastal water bodies along the South Atlantic Ocean. It has been pointed as a promising fish species to monitor the health condition of tropical and sub-temperate coastal waters. Objective: The present study evaluated biochemical parameters in guppy exposed to waterborne Zn with or without salt water (24 ppt), in order to verify protective effects of salinity. Activity of catalase (CAT), total peroxides (H 2 O 2 ) and lipid peroxidation (TBARS) were evaluated in whole body after 96h of exposure to Zn (0.5 mg/L). Methods: Fish were divided in four groups: CT (tap water), Zn (0.5 mg/L), Salt (24 ppt) and Salt (24 ppt) + Zn (0.5 mg/L) during 96h. After, were anesthetized and euthanized by punching the spinal cord behind the opercula. Whole body was prepared though homogenization in Tris- HCl 50 mM, pH 7.5 for biochemical analyzes The protocols were previously approved by the Ethics Commission on Animal Use of the Federal University of Santa Maria under process number 117/2013. Results: CAT activity increases the Salt group and Salt + Zn compared to the CT and Zn groups. The group exposed to Zn increase the amount of total peroxides. However, in salt water groups, the levels remained similar to CT. Lipid peroxidation increased in Zn and Salt groups. Comparison between groups showed that TBARS in group Sal+Zn did not differ to CT, but decreased as compared to Zn and Salt groups. Conclusion:

These results suggest that salt in water per se increased the CAT activity and showed clear relation with peroxides reduction observed in the same groups. Zinc group increased peroxides formation, and this occurrence is prevented by salt in water. The association between salt and zinc prevent lipid peroxidation induced by zinc and salt per se. In fact, the competition of zinc and compounds of salt especially calcium could be the reason of positive results presented.

Financial Support: CAPES

Key words: fish; metal; salt water.

INTERESTERIFIED FAT CHRONIC SUPPLEMENTATION FACILITATES OXIDATIVE DAMAGE IN HIPPOCAMPUS OF RATS Lívia F. D’avila 1

INTERESTERIFIED FAT CHRONIC SUPPLEMENTATION FACILITATES OXIDATIVE DAMAGE IN HIPPOCAMPUS OF RATS

Lívia F. D’avila 1 , Verônica T. Dias 1 , Luciana T. Vey 2 , Karine Roversi 1 ,Laura H. Milanese 1 , Fabíola Trevizol 1 , Marilise E. Bürger 1,2

1 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria. 2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: Interesterification is one of the processes used to modify the physico-chemical characteristics of oils and fats, which involves redistribution of fatty acids (FA) between and within triacylglycerols molecules, until a thermodynamic equilibrium is reached. It has been used as an alternative for preparing plastic fats with low levels of trans isomers or absence of these FA. Studies involving interesterified fat (IF) have shown deleterious effects on the cardiovascular system, predisposition to obesity and increased incidence of type II diabetes, developing similar adverse effects related to hydrogenated vegetable fat consumption, however, there are no studies on the effect of its consumption on the central nervous system (CNS). Objective:

The aim of this study was to investigate whether the chronic consumption of IF may affect oxidative parameters in hippocampus of rats. Methods: To perform this trial, one week before mating, female adult Wistar rats (n=14) were supplemented (3g/kg; per oral) with either soybean oil, (SO-C, isocaloric control group) or interesterified fat (IF) and maintained under the same supplementation during pregnancy and lactation. From weaning, at the postnatal day (PND) 21, one male pup of each litter (n=7) was maintained on the same original oral supplementation until PND 90. Oxidative status of the hippocampus was estimated by reactive species (RS) generation, protein carbonyl (PC) levels and catalase (CAT) activity (process number 1391090616/2016). Results: IF supplemented group showed increased RS generation and PC levels and also decreased activity of CAT in hippocampus, in comparison to SO-C group. Conclusion: These data suggest that IF intake from pregnancy to adulthood is able to increase oxidative damages in hippocampus of adult offsprings. Considering that the increase of oxidative stress in CNS is related to neurodegenerative and neuropsychiatric diseases, additional studies should be carried out to evaluate the impact of IF consumption on the development of these diseases. Additional studies involving the influence of IF consumption over generations on neurochemical and epigenetic changes are required.

Financial Support: CAPES.

Key words: reactive species; first generation; central nervous system.

Uncaria tomentosa EXTRACTS MODULATES SPATIAL MEMORY BY PROMOTES ANTI-ACETYLCHOLINESTERASE ACTIVITY IN AGED RATS Lívia G.

Uncaria tomentosa EXTRACTS MODULATES SPATIAL MEMORY BY PROMOTES ANTI-ACETYLCHOLINESTERASE ACTIVITY IN AGED RATS

Lívia G. Castilhos 1,2 , Fatima H. Abdalla 2 , Jamile F. Gonçalves 2 , Fernanda L. Cabral 1 , Alessandra G. Manzoni 2 , Daniela B.R. Leal 1,2

1 Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal de Santa Maria.

2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: Senescence is the progressive physiological process of deterioration leading to a decline in fitness with age, which is not synonymous with infirmity and frailty associated with extreme old age in humans and animals. Uncaria tomentosa (Willd. ex Roem. & Schult.) DC., commonly known as cat’s claw have been reported to be neuroprotective via regeneration of the neuronal network promising activity in neuropsychopharmacology. Substantial evidence suggests that cholinergic neurotransmission regulates adult neurogenesis, and a depletion of cholinergic input to the neurogenic milieu during the aging process may contribute to spatial memory deficits. Objective: The aim of this study was to investigate if U. tomentosa improve cognition during senesce process through behavior tests and AChE activities on cerebral structures. Methods: Rats were treated with U. tomentosa (5mg/kg/ 15mg/kg, 30mg/kg) during one month and one year. All behavioral measures were assessed using the ANY-Maze TM software and the following endpoints were analyzed: scape latency and time spent in zones. The AChE enzymatic assay was determined by a method of Ellmann et al. (1961). The protocols were previously approved by the Ethics Commission on Animal Use of the Federal University of Santa Maria under process number 098/2014. Results: The escape latency from was significantly shorter for animals that received 5mg/kg, 15mg/kg and 30mg/kg of U. tomentosa than control, during 1 month or 1 year. The groups that received 5mg/kg, 15mg/kg and 30mg/kg of U. tomentosa prolonged swimming time in platform zone when compared to control. The results obtained for cerebral cortex AChE activity in senil animal treated with U. tomentosa for 1 month showed significant alteration. The doses of 15mg/kg and 30mg/kg decreased AChE activity. Besides that, the three doses of U. tomentosa reveled alteration on AChE activity. The doses of 5mg/kg, 15mg/kg and 30mg/kg reduced AChE activity for cerebral cortex (p<0.05), in comparison to control. Conclusion: These data suggest that U. tomentosa extracts are an interesting herb in terms of its potential use to improve cognitive function during senescence process.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: Uncaria tomentosa; cat’s claw; acetylcholinesterase; senescence.

TOXICITY OF AGROCHEMICAL MANCOZEB IN EMBRYO DEVELOPMENT AND BEHAVIOR OF ZEBRAFISH Luana Paganotto Leandro 1

TOXICITY OF AGROCHEMICAL MANCOZEB IN EMBRYO DEVELOPMENT AND BEHAVIOR OF ZEBRAFISH

Luana Paganotto Leandro 1 , Dennis Guilherme da Costa Silva 2 , Andressa Rubim Lopes 1 , Illana Kemmerich Martins 2 , Lucia Emanueli Schimith 3 , Nelson Rodrigues de Carvalho 2 , Mauro Eugênio Medina Nunes 4, Thais Posser 5 , Jeferson Luis Franco 5.

1 Ciências Biológicas Bacharelado, Universidade Federal do Pampa; São Gabriel, Rio Grande do Sul;

2 Programa de Pós-Graduação em Ciências Biológicas da Universidade Federal do Pampa; São Gabriel, Rio Grande do Sul;

3 Biotecnologia, Universidade Federal do Pampa; São Gabriel, Rio Grande do Sul;

4 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica da Universidade Federal de Santa Maria; Santa Maria, Rio Grande do Sul;

5 Universidade Federal do Pampa; São Gabriel, Rio Grande do Sul.

Introduction: Mancozeb (MZ), a manganese/zinc-containing dithiocarbamate fungicide, shows low acute toxicity and low environmental persistence, however, its degradation products have been shown to cause impaired embryogenesis in non-target organisms. Zebrafish, a model organism, has been used to characterize toxicological effects mediated by dithiocarbamates in the embryo-larval phase, but the molecular mechanisms involved in MZ embryotoxicity are not well understood. Objective: In this context, we aimed to investigate the involvement of oxidative stress in the embryotoxicity induced by MZ in zebrafish. Methods: The exposure method was performed according to OECD GUIDELINES FOR THE TESTING OF CHEMICALS 236 - Fish Embryo Acute Toxicity (FET) Test, 2003. Embryos were maintained in 24 well plate containing system water (pH 7.2, 500µ.S cm -1 , 28°C). Embryos, 3-4 hours post fertilization (hpf) were pre-exposed during 1h to the antioxidant N-acetylcysteine (NAC; 750μM). Subsequently, the embryos were exposed to MZ concentrations (0.5, 0.75 and 1.0 mg L -1 ). Controls were maintained in system water only. Embryo-larval development, mortality, and a number of hatched embryos were evaluated as embryotoxic end points. For the biochemical and behavioral analysis, MZ 0.5 mg L -1 was used since it did not cause severe morphological alterations. Reactive oxygen species (ROS) formation was evaluated as DCFH-DA oxidation assay. The sensory-motor activity was evaluated through of the touch scape test and exploratory behavior (open field) was measured using ANY-Maze TM software. Results: MZ, at all concentrations tested induced dose-dependent morphological abnormalities, in addition to a low preference to the periphery and decreased distance traveled in the sensory- motor activity test. In parallel, MZ-treated embryos showed decreased speed and distance traveled in the open field apparatus. It was also observed a significant increase in ROS formation in MZ-treated embryos. Pre- treatment with NAC blocked all morphological changes, locomotor /sensory responses and decreased MZ- induced ROS. Conclusion: It was shown that MZ induces developmental and sensory-motor deficits accompanied by ROS formation in zebrafish embryos. Since a potent antioxidant (NAC) was able to block MZ-induced deleterious effects, our results point to oxidative stress as an important mechanism involved in MZ embryotoxicity.

Financial Support: CAPES, CNPq, FAPERGS, UNIPAMPA.

Key Words: dithiocarbamates; embryotoxicity; ROS.

ACUTE EXPOSURE TO PERMETHRIN INDUCES LOCOMOTIVE DEFICIT AND OXIDATIVE DAMAGE IN LARVAL ZEBRAFISH Lucia E.

ACUTE EXPOSURE TO PERMETHRIN INDUCES LOCOMOTIVE DEFICIT AND OXIDATIVE DAMAGE IN LARVAL ZEBRAFISH

Lucia E. Schimith 1 , Mauro E.M. Nunes 2 , Dennis G.C. Silva 3 , Nelson Rodrigues de Carvalho 3 , Andressa R. Lopes 4 , Luana P. Leandro 4 , Thais Posser 3 , Jeferson L. Franco 5

1 Biotecnologia Bacharelado, Universidade Federal do Pampa. São Gabriel, Rio Grande do Sul.

2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica da Universidade Federal de Santa Maria, Santa Maria, Rio Grande do Sul.

3 Programa de Pós-Graduação em Ciências Biológicas da Universidade Federal do Pampa. São Gabriel, Rio Grande do Sul.

4 Ciências Biológicas Bacharelado, Universidade Federal do Pampa. São Gabriel, Rio Grande do Sul.

5 Orientador. Universidade Federal do Pampa.

Introduction: Pyrethroid insecticides, such as permethrin (PM), are one of the most commonly used insecticides, due to its low toxicity to non-target organisms. However, several studies suggest that developing animals are uniquely susceptible to pyrethroid toxicity. Zebrafish have recently been used as an alternative model to assess the developmental neurotoxicology effects, due to small size, easy husbandry, high fecundity and its genetic and physiological similarity to mammals. Objective: The aim of this study was to investigate the behavioral effects and its influence on oxidative stress biomarkers in larval zebrafish exposed to PM. Methods: Larval zebrafish at 6-day postfertilization (dpf) were exposed to 25 - 600 µg.L -1 of PM diluted in ethanol during 24h. The final concentration of ethanol in each treatment was 0.01%. Controls (CTL) were treated with 0.01% ethanol. Locomotor and exploratory activities were analyzed in the novel tank test, which may reflect habituation to novelty stress. All behavioral measures were assessed using the ANY-Maze TM software and the following endpoints were analyzed: distance traveled; number of entries and time to the center of arena. For oxidative stress biomarkers, 50 larvae were compiled (one sample = fifty pooled larvae). Lipid peroxidation was estimated by thiobarbituric acid reactive substances (TBARS) production, catalase (CAT), glutathione peroxidase (GPx) and glutathione S-transferase (GST) activities were used as biomarkers of oxidative stress. Results: Firstly, we evaluated the cumulative mortality (LC50=37 µg.L -1 ). After, two concentrations were chosen, one lower and one higher than LC50 (25 and 50 µg.L -1 ). Both exposed groups showed significant decrease in distance traveled and thigmotaxis when compared to CTL. Thigmotaxis was observed by significant increase in permanence in the central arena. In parallel to the behavioral changes, PM treated larvae showed oxidative damage observed by significant increases in TBARS levels and GST activity. CAT and GPx activities were not changed. The increase in GST activity may be related to detoxification of xenobiotic and elimination of unsaturated aldehydes produced during the lipid peroxidation process. Conclusion: These data suggest that zebrafish larvae are sensitive to PM exposure, which causes mortality and behavioral deficits, as well as accumulation of oxidative damage, highlighting its potential toxicity during developmental stages.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: pyrethroid toxicity; developmental stages; permethrin; behavioral changes; oxidative stress.

PROTECTIVE EFFECT OF DIPHENYL DISELENIDE IN ZEBRAFISH SUBJECTED TO HYPOXIA Marcos M. Braga 1 ,

PROTECTIVE EFFECT OF DIPHENYL DISELENIDE IN ZEBRAFISH SUBJECTED TO HYPOXIA

Marcos M. Braga 1 , Ben Hur M. Mussulini 2 , Emerson S. Silva 2 , Gabriela Lazzarotto 2 , Diogo L. Oliveira 2 , Diogo O. Souza 2 , João Batista T. Rocha 1

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica, Universidade Federal do Rio Grande do Sul.

Introduction: Hypoxia is a condition found commonly in several disorders, such as ischemia. Since the brain is known for elevated basal O 2 consumption, the mitochondrial activity in this organ is promptly affected by hypoxia. A preventive or reverse treatment focusing on mitochondrial dysfunction induced by brain hypoxia could be obtained by diphenyl diselenide (PhSe) 2 , an organoselenium compound containing elevated antioxidant capacity. Thus, the use of the recently developed hypoxia model becomes appropriate to evaluate the effects of (PhSe) 2 in oxygen deprivation. Objective: The aim of this study was to investigate whether (PhSe) 2 prevent and/or revert behavioral and mitochondrial impairments performed hypoxia in adult zebrafish. Methods: Based on previous dose screening, animals were administered with a single i.p. injection of 100 mg/kg (PhSe) 2 dissolved in soybean oil. To evaluate the preventive effect (pre-treatment) of (PhSe) 2 , the compound was administered 3h prior to hypoxia trial. Fish subjected to pre-treatment were observed during hypoxia in order to obtain the latency time to reach the 3 rd behavioral stage of hypoxia. To investigate the reverse effect (post-treatment) of (PhSe) 2 , the compound was administered immediately after hypoxia and, after 1h, animals were evaluated in novel tank test, while their brain were separated to quantify the tissue amount of (PhSe) 2 by HPLC technique and to analyze the mitochondrial activity by TTC staining and high resolution respirometry (HRR). The protocols were previously approved by the Ethics Commission on Animal Use of the Federal University of Santa Maria (number 2523130115). Results: The compound did increase the resistance to hypoxia as observed in pre-treated animals. Post-treatment with (PhSe) 2 mitigated mildly the behavioral changes induced by hypoxia. However, this same treatment did significantly recover mitochondrial activity (e.g., ATP production). Conclusion: These data showed that (PhSe) 2 exerts a protective effect on hypoxia, which is strongly associated to a reverse action on mitochondrial dysfunction induced by hypoxia.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: diphenyl diselenide; hypoxia; zebrafish.

THE TOXICITY OF ANATOXIN-A (S) ON NEUROBIOLOGICAL PREPARATIONS OF BIRD, MAMMAL AND INSECT Maria E.

THE TOXICITY OF ANATOXIN-A (S) ON NEUROBIOLOGICAL PREPARATIONS OF BIRD, MAMMAL AND INSECT

Maria E. Rosa 1 , Douglas S. dos Santos 3 , Ana P. Zanatta 1 , Bruna T. Borges 1 , Barbara A. B. Ogata 1 , Lucas Maciel 1 , Yuri C. Barreto 1 , Allan P. Leal 1 , Etiely Karnopp 1 , Lúcia Vinadé 1 , Ernani Pinto 2 , Cháriston A. Dal Belo 1,3

1 Laboratório de Neurobiologia e Toxinologia (LANETOX), Universidade Federal do Pampa. 2 Departamento de Análises Clínicas Toxicológicas da Faculdade de Farmácia, Universidade de São Paulo. 3 Programa de Pós-graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: The anatoxin-a(s) is a potential biohazard cyanotoxin, in which toxic mechanism relies on the inhibition of the acetylcholinesterase (AChE) enzyme. Despite the ecological and clinical interest, there are feel works in literature aimed to study the interactions of anatoxin-a(s) on the nervous system of different animal species. Objective: In this study we investigated the neurotoxic activity induced by the extract containing anatoxin-a(s) (ECATAS) on in vivo and in vitro neurobiological preparations of invertebrate and vertebrates. Methods: The extract containing anatoxin-a(s) was a donation of Dr. Ernani Pinto, from University of São Paulo, USP. Adult male Nauphoeta cinerea (Olivier) cockroaches (34 months after adult molt), Swiss white mice (25-30g) and Hy-Line chicks (1-10 days) were used for the biological assays. The animals were kept with water and food ad libitum in controlled temperature and lighting environment. The in vitro chick biventer cervicis muscle preparation (BCMP) was used for vertebrate neuromuscular assessment. The in vivo cockroach neuromuscular preparation (CNP), was mounted for the neuromuscular assays in insects, as described elsewhere (Martinelli et al. 2014, Biochimica et Biophysica Acta 1840, 935944). The cell viability of brain slices was accessed using the MTT assay. Acetylcholinesterase (AChE) activity was accessed using cockroach brain homogenates in a conventional methodology. Data were expressed as mean ± standard error (SE) and analyzed by Student “t” test or ANOVA followed by Tukey test. This work was approved by the Institutional Committee for Ethics in Animal Use (CEUA/ UNIPAMPA, Protocol nº 043/2015). Results:

ECATAS (2.5, 5 and 50µg/ml, induced a significative inhibition of the AChE, that was maximal (53.39 ± 1.65%, n= 6, p<0.0001), for the highest concentration. The cell viability of mice hippocampal slices showed that the ECATAS (1, 4 and 8μg/ml) induced a maximum decreased in the cell viability (4μg/ml), compared to the HEPES control (p <0.05, n=6). Using BCMP and CNP, ECATAS (2, 4 and 8µg/ml) and (5, 25 and 50 µg/g of animal weight), produced a transient increase of twitch tension (20± 1.65% and 10± 1%) followed by an irreversible neuromuscular blockade that was maximum and complete for the lowest concentration (p<0.01, n=6, respectively). Conclusion: The data indicated that different species are susceptible to the toxic effect of the anatoxina-a(s). Also that the toxin may affect both peripheral and central nervous systems. This work unveil the use of neurobiological preparations as biomarkers, while shows the biotechnological potential of anatoxins.

Financial Support: Edital Toxinologia 063/2010 CAPES, Edital Grupos de Pesquisa UNIPAMPA 2014.

Key words: cyanotoxins; anti-acetylcholinesterase; neuromuscular junctions; central nervous system.

EFFECTS OF N-ACETYL-CYSTEINE AND ZINC IN BRAIN CHANGES CAUSED BY LEADEXPOSURE IN YOUNG RATS Mariana

EFFECTS OF N-ACETYL-CYSTEINE AND ZINC IN BRAIN CHANGES CAUSED BY LEADEXPOSURE IN YOUNG RATS

Mariana S. C. dos Santos 1,2 , Taíse F. Pedroso 2 , Maria Ester Pereira 2,3

1 Centro de Ciências da Saúde (CCS)-Federal University of Santa Maria, Santa Maria, RS, Brazil. 2 Programa graduate program in life sciences: Toxicological Biochemistry, Federal University of Santa Maria, Santa Maria, RS, Brazil. 3 Departamento of Biochemistry and Molecular Biology, Natural and exact sciences(CCNE)-Federal University of Santa Maria, Santa Maria, RS, Brazil.

Introduction: Lead (Pb) is a toxic metal, which has no biological function, can cause several adverse reactions in individuals exposed to him, the most serious of them is due to the accumulation of Pb in the brain. The acetylcholinesterase (AChE) is an enzyme that breaks down acetylcholine, neurotransmitter found in the brain which is responsible, among others, by nerve impulses, when in excess cause about muscle stimulation and cases of dementia, when accumulation of acetylcholine is the brain. Knowing that young rats have greater sensitivity to external aggressions, it is believed that a Pb exposure in the early days postnatal would cause biochemical damage and behavioral changes. Objective: Today, it is known that the treatment of metal poisoning is done with chelating agents, which can cause adverse effects such as the depletion of essential metals, that way, the search for alternative treatment is important. Zinc (Zn) and N-acetilcisteina (NAC) are compounds that possess important properties against the toxic effects of metals, which may have a protective effect against Pb poisoning search so, investigate the possible protective action of ZnCl 2 and the NAC on the toxicity of Ac 2 PbWistar rats. Methods: The animals were treated as follows: received saline, zinc chloride (ZnCl 2 27 mg/kg), N-acetylcysteine (NAC 5 mg/kg) or ZnCl 2 + NAC day 3º to 7º; lead acetate (Ac 2 Pb 1 th to 12 th days of age by subcutaneous route. The animals were subjected to the tests of geotact negative and open field. The 33 days of age, were euthanized and removed brain and cerebellum for measurement of the activity. Results: the chicks exposed to Ac 2 Pb show decrease in the activity of the enzyme AChE, but there were no behavioral changes in geotact x negative test and open field while both preventative care (Zn, ZN + NAC and NAC) protected partly inhibition of AChE caused by Pb.

Financial Support: CAPES, CNPq and PRAE/UFSM.

Key words: lead; acetylcholinesterase; neurotoxicity.

Paullinia cupana AND CAFFEINE REDUCED THE TRIACILGLICEROL CONTENT OF Caenorhabditis elegans SUPPLEMENTED WITH GLUCOSE

Paullinia cupana AND CAFFEINE REDUCED THE TRIACILGLICEROL CONTENT OF Caenorhabditis elegans SUPPLEMENTED WITH GLUCOSE

Marina Lopes Machado¹, Letícia Priscilla Arantes¹, Thayanara Cruz da Silva¹, Tássia Limana da Silveira¹, Daniela de Freitas Câmara¹, Fabiane Bicca Obetine Baptista¹, Félix Alexandre Antunes Soares¹

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: Obesity is characterized by an excess of adipose tissue, and is associated with health problems and increased mortality. Many methods are used to treat obesity, most of them are pharmaceuticals, which might cause collateral effects. Paullinia cupana (guarana) is widely consumed in different regions of Brazil and has a high concentration of caffeine, which is known for its stimulant and thermogenic properties in vitro. Objective: The aim of this study was to investigate the effect of chronic exposure of Paullinia cupana or caffeine in vivo on triglyceride levels of Caenorhabditis elegans supplemented with glucose. Methods: The N2 wild-type or VS20 worms were treated from L1 stage to young-adult with Paullinia cupana at 1 mg/mL or caffeine at 50 uM. To perform the assay of worms with more lipid accumulation, 15 mM of glucose was added to the medium together with the treatments. The parameters analyzed were antimicrobial activity of the compounds, quantification of triglycerides and expression of adipose triglyceride lipase 1 (ATGL-1). Results:

The compounds analyzed did not present antimicrobial activity. Paullinia cupana or caffeine did not alter the triglycerides content of N2 worms. When the worms were treated with 15 mM glucose, the triglyceride levels were increased about 94% compared with control group, and the treatment with Paullinia cupana or caffeine plus glucose showed an increase in triglycerides about 40%, and this result is dependent on ATGL-1 expression. Conclusion: It was observed that neither Paullinia cupana nor caffeine presented any effect on baseline triglyceride levels, but when the animals were treated with glucose, caffeine and Paullinia cupana prevented, at least in part, the increase of triglyceride levels.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: triglycerides; obesity; guarana.

BIOLOGICAL POTENTIAL IN THE FAMILY Eriocaulaceae Micaela Ferreira Viana 1 , Andressa Palharini Machado 1

BIOLOGICAL POTENTIAL IN THE FAMILY Eriocaulaceae

Micaela Ferreira Viana 1 , Andressa Palharini Machado 1 , Mara Lisiane Tissot Squalli Houssaini 2 , Ilaine Teresinha Seibel Gehrke 2

1 Graduação em Ciências Biológicas- Bacharelado, Universidade Regional do Noroeste do Estado do Rio Grande do Sul.

2 Quadro docente do departamento de Ciências da Vida, Universidade Regional do Noroeste do Estado do Rio Grande do Sul.

Introduction: The elucidation of the active compounds presents in plants, as well as your action mechanisms, have been a big challenge to pharmaceutic chemistry, biochemistry and pharmacology. Eriocaulaceae is easily distinguished from other families by having inflorescences in the form of chapters. This family has about 1200 species distributed in 10 genres and is divides in two subfamilies: Eriocauloidae Ruhland e Paepalanthoideae Ruhland. The biological activity this Family started with the discovery of the metabolite paepalantine, classified as one in naphtophyran, that usually is not founded in plants but in fungal microorganisms like that of the genre Penicillium spp. The phytochemical and biological study together of this family is relevant to contribute with the description of bioactivities compunds, etnopharmacology, pharmacology and even ecology of this plant. Objective: The objective this study is seek to understand the mains results about the biological activity in the Eriocaulaceae family. Methods: This bibliographic research was related with articles selected of the research platform Pub Med and that they brought in your results biological effects caused for Eriocaulaceaes species. Results: Were found 27 articles that it is about the family Eriocaulaceae and of these just 12 works results on biological tests. These tests were positive therapeutic potential for Eriocaulaceae species, of which we can cite antifungal and antimicrobial activity mainly vulvovaginal candidiasis, in addition curative effect and gastroprotective activity. Another property important is the antimutagenic, antioxidant and anti-inflammatory potential. Moreover, specimens from this family may also be inhibitors of Aurora B Kinase. Conclusion: In this sense, the Eriocaulaceae family has relatively few and recent researches related to their biological potential and endemic species of Rio Grande do Sul do not have these characteristics still clarified yet. Therefore, the phytochemical and toxicological study of this family can be of great relevance in the search for new therapeutic alternatives.

Key words: Eriocaulaceae; review; activity biological.

BIOCHEMICAL ALTERATIONS ON CEREBRAL CORTEX AFTER ACUTE EXPOSURE TO AFLATOXIN B1 ON YOUNG RATS Naiéli

BIOCHEMICAL ALTERATIONS ON CEREBRAL CORTEX AFTER ACUTE EXPOSURE TO AFLATOXIN B1 ON YOUNG RATS

Naiéli Schiefelbein Souto 1 , Micheli Dassi 1 , Ana Claudia Monteiro Braga 2, Érica Furlan da Rosa 2 , Ana Flávia Furian 1,2

1 Programa de Pós-Graduação em Ciência e Tecnologia em Alimentos, Universidade Federal de Santa Maria.

2 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria

Introduction: Aflatoxins are mainly produced by Aspergillus flavus and A. parasiticus. Aflatoxin B1 (AFB1) is the most common mycotoxin and highly toxic, causing carcinogenic, mutagenic and teratogenic effects. This mycotoxin has been detected in important crops worldwide, including corn, peanuts, beans, rice, wheat, fruits and also in animal feed. The AFB1 is converted in the liver into 8,9-epoxide, a metabolite which reacts with

Many researches clarified the peripheral effects of the exposition to AFB1, however,

there are few studies explaining their effects on the central nervous system. Objective: The aim of our study was to evaluate the acute effects caused by oral administration of AFB1 on biochemical parameters of cerebral cortex. Methods: Young male Wistar rats received a single administration of AFB1 (250 µg/kg/i.g.) and 48 hours thereafter biochemical parameters were measured in the cerebral cortex. Antioxidant enzymes superoxide dismutase (SOD) and glutathione S-transferase (GST), protein carbonyls and levels of 3- nitrotyrosine, determination of ascorbic acid and non-protein sulfhydryl groups (NPSH) and lipoperoxidation, (TBARS) as well as changes in immunoreactivity of protein kinase A (PKA-Ser96) and protein kinase C (PKC- Ser957). The protocols were previously approved by the Ethics Commission on Animal Use of the Federal University of Santa Maria under process number 093/2014. Results: Acute intoxication by AFB1 causes neurotoxic effects, evidenced by a significant reduction in the levels of ascorbic acid and non-protein sulfhydryl groups, accompanied by the increase in immunoreactivity ratio of protein kinase C phosphorylated/total (p-PKC α Ser957/PKC α). Conclusion: AFB1 was able to cause neurochemical alterations. These results reinforce that monitoring mycotoxins levels in food are essential to guarantee food security.

proteins, RNA and DNA.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: mycotoxin; biochemical parameters; neurotoxic.

In silico STUDIES OF INTERACTIONS BETWEEN RESVERATROL ISOMERS AND THE ENZYMES MAO-A/B Pablo A. Nogara

In silico STUDIES OF INTERACTIONS BETWEEN RESVERATROL ISOMERS AND THE ENZYMES MAO-A/B

Pablo A. Nogara 1 , Alcindo Busanello 2 , Nilda B. V. Barbosa 1 , Roselei Fachinetto 1,2 , João B. T. Rocha 1

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria. 2 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria, RS, Brazil.

Introduction: Monoamine oxidases (MAO) are flavoenzymes that catalyze the oxidative deamination of neurotransmitters (serotonin, dopamine, and norepinephrine). Based on the MAOs role, for instance, inactivation of neurotransmitters, selective inhibitors of MAO-A are used as antidepressants; whereas selective MAO-B inhibitors could be used in the treatment of Alzheimer's disease and Parkinson's disease. In this context, resveratrol (natural phenolic compound, which exists as cis and trans isomers) could be a MAO inhibitor, since it presented an antidepressant-like effect in animal. The molecular docking is a method which predicts the binding-conformation of small molecules in a protein target. Thus, the characterization of the binding behavior plays an important role in rational design of drugs as well as to elucidate fundamental biochemical processes. Objective: In this study, we propose evaluated the molecular interactions and the binding free energy (∆G) between resveratrol isomers and the enzymes MAO-A/B using molecular docking method. Methods: The program AutoDock Vina 1.1.1 was used in the blinding molecular docking. The crystal structure of MAO-A from rat (rMAO-A), was obtained from Protein DataBank (1o5w) and the rMAO-B was generated by protein homology modeling, using the web server Swiss-model, and the hMAO-B (2c73) as template. The cis and trans resveratrol molecules was created using the software Avogadro 1.1.1. Results:

According with the docking, resveratrol isomers have a great affinity by the active site of rMAO-A, with the m-benzenediol group close to the FAD, making a π-π stacking with Tyr407 and H-bonds with Asn181 and Tyr444. The analysis of ∆G, indicates that the trans-resveratrol (-9.1 kcal/mol) present a favorable binding to rMAO-A, than cis-resveratrol (-8.7 kcal/mol). On the other hand, the binding of resveratrol isomers with rMAO-B showed that the trans-resveratrol interacts in the active site, by H-bonds with Cys172 and Tyr345, while the cis isomer binds in another site, close to the transmembrane helix by π-π stacking with Phe103 and H-bonds with Asn116 and Asp123. The ∆G indicated that both trans/cis-resveratrol interact with rMAO-B with similar binding energy (-8.2 and -8.1 kcal/mol, respectively), however, this binding is less favorable, when compared with rMAO-A. Conclusion: These findings suggest that the trans-resveratrol presents more affinity to the rMAO-A, when compared with cis isomer and the rMAO-B.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: monoamine oxidases; resveratrol; molecular docking.

ISOFLAVONES PREVENT LIPID PEROXIDATION INDUCED BY PRO- OXIDANTS. Renata G. Budel 1 , Larissa F.

ISOFLAVONES PREVENT LIPID PEROXIDATION INDUCED BY PRO- OXIDANTS.

Renata G. Budel 1 , Larissa F. Schaffer 2 , Roselei Fachinetto 3 , Luis R. Peroza 2

1 Programa de Pós-Graduação em Nanociências, Centro Universitário Franciscano.

2 Docentes do curso de Biomedicina, Centro Universitário Franciscano.

3 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria.

Introduction: Oxidative stress is a biological condition that occurs due to an imbalance between antioxidant defenses and reactive oxygen species and/or reactive nitrogen species generated from normal oxidative metabolism or from pro-oxidant environmental exposures. This condition has been described to be involved in neurodegenerative disorders, where ROS can contribute to their worsening by causing alterations in the cell membrane and DNA mutations. It is known that the brain is vulnerable to oxidative damage because of a relative lack of antioxidant enzymes like catalase and glutathione peroxidase and abundance of oxidizable substrates like polyunsaturated fatty acids and catecholamines such as dopamine. Literature data show that genistein, one isoflavone, may be a promising agent for the treatment of neurodegenerative diseases, due to its ability to cross the blood brain barrier, long half-life (15-22 h) and low oral toxicity in vivo. Objective: This study was designed to investigate the potential protective effect of isoflavones on the damage of brain oxidative stress induced in vitro in brain tissue of rats. Methods: The isoflavones were weighted and dissolved in distilled water. For each experiment, a new solution was prepared. The brain samples were from male Wistar rats beheaded with the guillotine aid after anesthesia with Ketamine/Xylazine. To evaluate the effects of isoflavones on TBARS production induced by different pro-oxidants in vitro, the rat brain tissue was removed and homogenized. After, an aliquot of the supernatant was incubated for 1 hour at 37°C with pro-oxidants and in the presence or absence of different concentrations of isoflavones (equivalent to 25, 50, 100, 200 and 300 µg/mL of isoflavones). As pro-oxidant agents, sodium nitroprusside (NPS 5μM) and Fe 2+ /EDTA (100 μM/100 μM) were used. Malondialdehyde was used as standard and samples were read at 532 nm. All experiments were performed in accordance with the guidelines of the National Council of Control of Animal Experimentation (CONCEA). Results: Both NPS and Fe 2+ /EDTA complex increased TBARS in brain homogenates of the rat. Isoflavones reduced this increase in a concentration-dependent manner with an IC 50 of 71.39 ± 2.01μg/ml to NPS and 124.5 ± 8.13 μg/ml to Fe 2+ /EDTA. Conclusion: Here, the results show that isoflavones were able to reduce brain lipid peroxidation. One hypothesis to explain the effect of isoflavones on the lipid peroxidation induced by pro-oxidant agents is that isoflavones may have neutralized the inducing agents, thus avoiding the lipid damage.

Financial Support: CAPES.

Key words: TBARS; oxidative stress; isoflavones.

COGNITIVE AND CHOLINERGIC APPRAISEMENT OF MICE EXPOSED TO THE DIAZINON FOR 10 DAYS Sabrina Souza

COGNITIVE AND CHOLINERGIC APPRAISEMENT OF MICE EXPOSED TO THE DIAZINON FOR 10 DAYS

Sabrina Souza dos Santos 1,3 , Jamile S. Bernardi 2 , Maria Ester Pereira 2,3 .

1 Centro de Ciências da Saúde (CCS) - UFSM Universidade Federal de Santa Maria, Santa Maria, RS, Brasil. 2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria, Santa Maria, RS, Brasil. 3 Departamento de Bioquímica e Biologia Molecular, Centro de Ciências Naturais e Exatas (CCNE).

Introduction: Diazinon (DZ) is a pesticide belonging to the class of organophosphates widely used in agriculture and veterinary medicine. Its main toxic effect is the inhibition of cholinesterases (ChEs) - acetylcholinesterase (AChE, EC: 3.1.1.7) and butyrylcholinesterase (BChE , EC: 3.1.1.8). This inhibition causes the increase of the neurotransmitter acetylcholine (ACh) causing a greater stimulation of the cholinergic synapses. Objective: To assess the cognitive ability and cholinergic activity of mice exposed to DZ for a short period of time. Methods: This work was approved by the Ethics Committee on the Use of Animals of UFSM (CEUA / UFSM) under the number 6836140815. Twenty-three male albino mice of the Swiss strain were used, which were separated into 3 treatment groups (n = 7 to 8) and treated with saline (shan group) or DZ (10 or 100 mg / kg) for 10 days via the subcutaneous route. The open field (OF) test were performed on the 7 th day of exposure and the Morris Water Maze (MWM) test which was performed from the 7 th to 10 th days of exposure. Afterwards, the animals were anesthetized and euthanized to measure ChEs activity in the cerebral cortex and hippocampus. The results were considered significant when p<0.05. Results: Animals exposed to DZ showed no alterations in the OF task when compared to the shan. In the task of MWM, all the groups presented improvement of the performance with the passage of the days days=11.68; p<0.001], but there was no difference between the groups. DZ also did not alter the activity of ChEs in the cerebral cortex and hippocampus. Conclusion: DZ did not cause behavioral changes and did not alter the activity of brain ChEs. It is suggested to be related to the short exposure period.

Financial Support: CAPES, CNPq e PRAE/UFSM.

Key words: Organophosphate; memory; neurotoxicity; diazinon.

COGNITIVE AND CHOLINERGIC APPRAISEMENT OF MICE EXPOSED TO THE DIAZINON FOR30 DAYS Sara de Lima

COGNITIVE AND CHOLINERGIC APPRAISEMENT OF MICE EXPOSED TO THE DIAZINON FOR30 DAYS

Sara de Lima Marion 1,2 , Jamile S. Bernardi 2 , Maria Ester Pereira 2,3 .

1 Centro de Ciências da Saúde (CCS) - UFSM Universidade Federal de Santa Maria, Santa Maria, RS, Brasil. 2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria, Santa Maria, RS, Brasil. 3 Departamento de Bioquímica e Biologia Molecular, Centro de Ciências Naturais e Exatas (CCNE).

Introduction: Organophosphorus compounds (OP) form a large class of commonly used pesticides. Diazinon (DZ) is an example of this class and, as with other OFs, its main toxic effect is the inhibition of cholinesterases (ChEs) acetylcholinesterase (AChE, EC:3.1.1.7) and butyrylcholinesterase (BChE, EC:3.1.1.8). This inhibition results in the accumulation of the neurotransmitter acetylcholine (ACh) causing a greater stimulation of the cholinergic synapses. Objective: To evaluate the effects of prolonged exposure with two different doses of OF Diazinon, andalterations in spatial memory and levels of ChEs. Methods: This work was approved by the Ethics Committee of UFSM, number 6836140815. Twenty male Swiss albino mice were divided into three experimental groups (n= 4-9): Shangroup (saline), Group DZ 10 (10 mg/kg Diazinon); and Group DZ 100 (100 mg/kg Diazinon), by 30 days, via subcutaneous rout.On the 27 th day of exposure, the open field test (OF) was performed according to Walsh and Commins (1976), in order to evaluate the spontaneous locomotor activity. From the 27 th to the 30 th day of exposure, the Morris Water Maze (MWM) task was performed according to the method of MORRIS (1984), for the evaluation of spatial memory and learning. Afterwards, the animals were anesthetized and euthanized to measurement ofthe enzymatic activity of ChEs in cerebral cortex and hippocampus. Results: The results show that DZ did not alter the locomotor activity and the learning capacity of the animals. In the OP task, the animals exposed to DZ 10 showed a significant difference

in the time of exit [T (13) =12.234, p=0.044] and in the number of fecal cakes [T (13) =7.840, p=0.018]compared to the shan. In the MWM task, all groups showed improvement over the days [F (3,54) =35.83, p<0.001] and the animals exposed to DZ 10 presented lower latency to find the platform on the first day of the test [F (2,18) =4.231,

p =0.31] compared to the shan. The animals exposed to DZ 100 had a reduction in hippocampal AChE activity

[F (2,16) =3.779, p=0.045] and BChE of the cerebral cortex [F (2,17) =5.989, p=0.014 ],compared to the shan.

Conclusion: The locomotor activity and learning ability of animals exposed to DZ was not altered, as seen in behavioral tests. However, a higher dose altered BChE activity of the cerebral cortex and hippocampus AChE.

It is suggested that such changes in ChEs activity are not sufficient to cause behavioral damage.

Financial Support: CAPES, CNPq e PRAE/UFSM.

Key words: Organophosphate; memory; neurotoxicity; diazinon.

SODIUM SELENITE PREVENTS PARAQUAT-INDUCED NEUROTOXICITY IN ZEBRAFISH: BIOCHEMICAL EVIDENCE Talise Ellwanger Müller 1

SODIUM SELENITE PREVENTS PARAQUAT-INDUCED NEUROTOXICITY IN ZEBRAFISH: BIOCHEMICAL EVIDENCE

Talise Ellwanger Müller 1 , Mauro Medina Nunes 1 , Charlene Cavalheiro Menezes 1 , Flavia Vestena Stefanello 1 , Vania Lucia Loro 1 , Denis Broock Rosemberg 1

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: Paraquat (PQ) is considered a potential environmental neurotoxin that has been associated with increased risk for neurodegenerative diseases, such as Parkinson’s disease. In animals, PQ acts mainly on dopaminergic neurons due to its specificity with the same transporter used by L-dopa and produces high levels of oxidative and nitrosative stress. These stress conditions induced by PQ redox cycle can alter cell membranes causing NADPH oxidation, impair the recycling of oxidized glutathione, and promote lipid peroxidation. In this context, antioxidants have emerged as potential therapeutics for the treatment of neurodegenerative diseases. Selenium (SE) is an essential micronutrient for maintaining normal brain function in vertebrates and its biological functions are mediated in part by selenoproteins. SE shows antioxidant and neuroprotective effects by scavenging free oxygen radicals, exerting a protective role on biomolecules, such as DNA, lipids, and proteins by reducing hydroperoxides and lipoperoxidases. Objective: To investigate the protective effect of Na 2 SeO 3 dietary on biochemical parameters of zebrafish exposed to PQ. Methods: Initially, adult zebrafish of both sexes were divided into two groups: control diet (without Na 2 SeO 3 ) and SE diet (1.0 mg/Kg Na 2 SeO 3 ) for 21 days. After, the groups were subdivided into four experimental groups (n=25 each): CT group [control diet + saline treatment], PQ group [control diet + PQ treatment], SE group [SE diet + saline treated], and SE+PQ group [SE diet + PQ treatment]. 20mg/kg PQ and 0.9 % saline solution were administrated intraperitoneally (16 days, totalized six injections). After the treatment period, the animals were euthanized and biochemical analyzes were performed. Results: Na 2 SeO 3 dietary prevented the increase of carbonylated protein (CP), reactive oxygen species (ROS) and nitrite/nitrate (NOx) levels, as well as the decrease in non- protein thiols (NPSH) levels caused by PQ. Regarding the antioxidant enzymatic defenses, Na 2 SeO 3 dietary prevented the increase in catalase (CAT) and glutathione peroxidase (GPx) activities induced by PQ. Conclusion: Altogether, Na 2 SeO 3 dietary improves biochemical function impaired by PQ treatment in zebrafish, by modulating redox parameters. This study also validates the use of zebrafish for the study of potential protective molecules in models of neurodegenerative diseases.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: Selenium; Zebrafish; Oxidative stress; Parkinson’s disease.

QUINOLIC ACID INDUCES TOXICITY IN Caenorhabditis elegans Tássia L. da Silveira¹, Daniele C. Zamberlan¹, Marina

QUINOLIC ACID INDUCES TOXICITY IN Caenorhabditis elegans

Tássia L. da Silveira¹, Daniele C. Zamberlan¹, Marina L. Machado¹, Thayanara C. Silva¹, Leticia P. Arantes¹ Fabiane B. O. Baptista¹, Felix A. A. Soares¹

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: 2,3-Pyridinedicarboxylic acid, widely known as quinolinic acid (QA), is a metabolite of tryptophan degradation in kynurenine pathway, which acts as a NMDA receptor agonist. Furthermore, QA has been described as a potent endogenous neurotoxin related to various psychiatric disorders and neurodegenerative processes. The nematode Caenorhabditis elegans has a nervous system highly conserved with mammals and thus is an alternative animal model widely used in neurobiology research. However, there is no neurotoxin described that allows the study of glutamatergic system injuries in C. elegans. Objective: The aim of this study was to validate the use of AQ to induce glutamatergic toxicity in C. elegans. Methods:

Nematodes from N2 (wild type), VM487 (nmr-1) and VC2623 (nmr-2) strains at the young adult stage were treated in liquid or agar containing QA (5, 10, 20, 50, 100 and 200 mM) or vehicle (M9 buffer) at 20 o C for 1 hour. The analyses included evaluation of QA’s effects on survival, behavioral parameters (pharynx pumping and locomotion), subcellular DAF-16 localization and reactive species generation after exposure to QA. Results: QA at high concentrations (50, 100 and 200 mM) induced mortality (~15%), while at low concentrations (10 and 20mM) altered the behavioral pattern of the nematodes. QA also activated DAF- 16/FOXO signaling pathway and increased reactive species levels compared to control in an nmr-1.- dependent manner. Conclusion: These data suggests that QA might be used for toxicological studies on glutamatergic system injuries in C. elegans.

Financial Support: CNPq, CAPES.

Key words: C. elegans; Quinolinic acid; Toxicity.

NEUROBIOLOGY OF Rhinella icterica (SPIX, 1824) TOAD VENOM IN in vitro PREPARATIONS OF VERTEBRATES Yuri

NEUROBIOLOGY OF Rhinella icterica (SPIX, 1824) TOAD VENOM IN in vitro PREPARATIONS OF VERTEBRATES

Yuri C. Barreto 1,2 , Alan P. Leal 1 , Raquel S. Oliveira 1 , Ana P. Zanatta 1 , Barbara A. B. Ogata 1 , Bruna T. Borges 1 , Etiely Karnopp 1,4 , Maria E. Rosa 1 , Lucas F. Maciel 1 , Douglas S. Santos 3 , Tiago G. dos Santos 2 , Cháriston A. Dal Belo 1,3 , Lucia Vinadé 1 .

1 Laboratório de Neurobiologia e Toxinologia (LANETOX), Universidade Federal do Pampa, São Gabriel, Brasil.

2

Laboratório de Estudos do Bioma Pampa (LEBIP), Universidade Federal do Pampa, São Gabriel, Brasil.

3 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: The venom of the Brazilian toad Rhinella icterica is potentially a rich source for bioactive molecules, although there is no previous work describing its biological activity. Objective:

The aim of this study was to investigate the neurobiological activity of R. icterica venom methanolic extract (MERIV) at central and peripheral nervous system of vertebrates. Methods: Adult male Swiss white mice (25-30g) and Hyline chicks (1-10 days) were used for the biological assays. The animals were kept with water and food ad libitum in controlled temperature and lighting environment. The in vitro biventer cervicis muscle preparation (BCMP) was used for neuromuscular assessment. The cell viability of brain slices was accessed using the MTT assay. Acetylcholinesterase (AChE) activity was accessed using biventer cervicis muscles in a conventional methodology. Data were expressed as mean ± standard error (SE) and analyzed by Student “t” test or ANOVA followed by Tukey test. This work was approved by the Institutional Committee for Ethics in Animal Use (CEUA/ UNIPAMPA,

Protocol nº 037/2012). Results:

When MERIV (5 and 10 µg/mL) was incubated on BCMP there was

an increase in the muscle twitches followed by an irreversible neuromuscular blockade, in 120min

recordings (60±15% and 46±6%, p<0.05, n=5, respectively).

 

MERIV (5, 10 and 20 µg/mL)

significantly

inhibited

AChE

activity

(23±5%,

56±5%

and

52±6%,

p<0.05,

n=3,

p<0.05,

respectively).

The incubation of muscles with digoxin (52 nM) or ouabain (0.2 nM), mimicked the

venom facilitatory and blocking activities (27±6% and 19 ± 4%, p<0.05, n=5, respectively). MERIV

increased the mouse brain cell viability at the lowest concentration (36±10%,

n=6, p<0.05), with no alteration with the other doses. Conclusion: MERIV interfere with the peripheral and central neurotransmission in vertebrates, probably by altering the cholinergic neurotransmission, confirming its biological potential.

(5, 10 and 20 µg/mL)

Financial

Support:

Edital

de

Toxinologia

CAPES

063/2010,

PRONEM

/FAPERGS/CNPq

003/2011.

Key words: Toad venom; Acetylcholinesterase; neurotoxicity.

FINAL PROGRAM ABSTRACTS SESSION: NEUROPSYCHOPHARMACOLOGY

FINAL PROGRAM

ABSTRACTS

FINAL PROGRAM ABSTRACTS SESSION: NEUROPSYCHOPHARMACOLOGY

SESSION: NEUROPSYCHOPHARMACOLOGY

ANESTHETIC ACTIVITY OF THYMOL AND CARVACROL IN SILVER CATFISH ( Rhamdia quelen ). Adriane E.

ANESTHETIC ACTIVITY OF THYMOL AND CARVACROL IN SILVER CATFISH (Rhamdia quelen).

Adriane E. Bianchini 1 , Jessyka A. da Cunha 1 , Isabel C. M. Brusque 2 , Guerino Bandeira Junior 2 , Bernardo Baldisserotto 1

1 Pharmacology Graduation Program, Universidade Federal de Santa Maria. 2 Veterinary Medicine course, Universidade Federal de Santa Maria.

Introduction: Anesthetics are used in aquaculture to improve animal welfare and natural products are good sources for the discovery of new drugs. The monoterpene thymol and carvacrol are present in several aromatic plants and their depressant activities in the central nervous system have been evidenced in rodents. Objective:

The aim of this study was to evaluate whether thymol and carvacrol have sedative and / or anesthetic effect in silver catfish (Rhamdia quelen). Methods: Fish (n = 6 / concentration and compound) (12.84 ± 0.34 g, 10.5 ± 0.22 cm) were exposed to 25, 50, 75 or 100 mg/L thymol or carvacrol, in a 1 L aquarium. The observation occurred until the animals reached deep anesthesia stage (fish does not respond to any external stimulus) or maximum time of 30 min. After, they were placed in anesthetic-free aquaria until complete recovery (total swimming capacity and response to stimulation with glass rod in the aquarium bottom) or maximum time of 30 min. Survival was assessed 48 h after the experiment. The protocol was approved by the Ethics and Animal Welfare Committee of the Federal University of Santa Maria (Process No. 074/2014). Results: Regression analysis showed a concentration-response relationship for thymol and carvacrol in all stages of anesthetic induction but not for recovery. Thymol and carvacrol were sedatives with 25 mg L -1 (611.00 ± 37.65 s and 352.33 ± 22.56 s, respectively) and carvacrol was significantly faster at induction. Deep anesthesia was produced with 50, 75 and 100 mg L -1 (thymol: 491.50 ± 10.49 s, 373.33 ± 15.85 s, 170.00 ± 11.87 s and carvacrol: 524.67 ± 36.09 s, 386.33 ± 9.26 s, 299.17 ± 13.42 s, respectively). Thymol 100 mg L -1 was significantly faster in inducing anesthesia. Recovery times were: 1649.00 ± 47.78 s, 919.67 ± 122.55 s and 1375.67 ± 150.86 s for 25, 50 and 75 mg L -1 thymol, respectively and 1580.83 ± 81.69 s for 50 mg L -1 carvacrol. The other concentrations had recovery time above 30 min. Carvacrol caused 50, 33, 33 and 16% of mortality at 25, 50, 75 and 100 mg L -1 , respectively. Involuntary muscular contractions were observed in animals exposed to carvacrol and to thymol, with less intensity. Conclusion: Thymol and carvacrol were effective in anesthetizing silver catfish, but the use of carvacrol for this purpose is not recommended because of the mortality induced after recovery.

Financial Support: CNPq and CAPES.

Key words: anesthetic; monoterpenes; fish.

ESSENTIAL OILS AS ANESTHETICS FOR Serrasalmus rhombeus Ana P. G. Almeida 1 , Berta M.

ESSENTIAL OILS AS ANESTHETICS FOR Serrasalmus rhombeus

Ana P. G. Almeida 1 , Berta M. Heinzmann 2 , Adalberto L. Val 3 , Bernardo Baldisserotto 1

1 Animal Biodiversity Graduation Program, Universidade Federal de Santa Maria.

2 Departmento of Industrial Pharmacy, Universidade Federal de Santa Maria.

3 Laboratório de Ecofisiologia e Evolução Molecular, Instituto Nacional de Pesquisas da Amazônia

Introduction: Practices performed in aquaculture as eggs extrusion, biometry and transport expose fish to intense stress, resulting in damage to the animal’s health. To minimize the stress effects anesthetics can be used to reduce fish motility and protect vital functions. However, synthetic anesthetics can cause adverse effects, so there is an interest in natural anesthetics with low toxicity and good economic viability, as essential oils (EO) extracted from plants. Objective: The aim of this study was to evaluate the anesthetic efficacy of EOs of Aloysia triphylla and Lippia alba in Serrasalmus rhombeus from the Rio Negro, Amazon (AM). Methods: Specimens of S. rhombeus (14. 9 ± 0. 51 cm) were collected during an expedition to Anavilhanas Islands of the Negro River, AM. Fish were transferred individually to aquaria containing 2L of water with the A. triphylla and L. alba EOs at 50, 100 or 150 µL L -1 and 50, 100 or 200 µL L -1 , respectively. The EOs were first diluted in ethanol at a proportion of 1:10. Time for anesthesia induction and recovery were evaluated. The maximum observation time was 15 min. Control experiments were performed using aquaria containing water and ethanol at a concentration equivalent to the highest dilution (1800 µL L -1 ). After induction of anesthesia, fish were transferred to a tank containing anesthetic-free water to evaluate the recovery time. Results: Fish exposed to 150 and 200 µL L -1 of A. triphylla and L. alba EOs, respectively, showed anesthetic induction time lower than 3 min and recovery time was lower than 10 min at all concentrations tested. Concentrations of 50 µL L -1 of both EOs caused only sedation. Conclusion: These results demonstrated that both EOs were effective for anesthesia of S. rhombeus.

Financial Support: CAPES, ADAPTA (CNPq-FAPEAM).

Key words: anesthesia; essential oils; recovery.

GABAPENTIN REDUCES HALOPERIDOL-INDUCED VACUOUS CHEWING MOVEMENTS IN MICE Ana Paula Chiapinotto Ceretta 1 , Catiuscia

GABAPENTIN REDUCES HALOPERIDOL-INDUCED VACUOUS CHEWING MOVEMENTS IN MICE

Ana Paula Chiapinotto Ceretta 1 , Catiuscia Molz de Freitas 2 , Larissa Finger Schaffer 1 , Mariana Maikéli Dotto 3 , Jeane Binotto Reinheimer 3 , Elizete de Moraes Reis 1 , Roselei Fachinetto 1,2

1 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria.

2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

3 Curso de Farmácia, Universidade Federal de Santa Maria, Santa Maria.

Introduction: Tardive dyskinesia is a common side effect in patients with long-term typical antipsychotic medication and vacuous chewing movement’s model induced by haloperidol has been used to study the abnormalities in animals. Its cause remains unclear, but several lines of evidence suggest that the dopamine receptor supersensitivity and gama-aminobutyric acid (GABA) insufficiency play an important role in the development of the disorder. Objective: The aim of this study was to investigate the effects of treatment of the GABA-mimetic drug gabapentin on the development of haloperidol-induced vacuous chewing movements (VCMs). Methods: Male mice received the vehicle, haloperidol (1.5 mg/kg) or gabapentin (GBP, 100 mg/kg) intraperitoneally during 28 consecutive days. Vacuous chewing movements (VCMs) quantification was evaluated before the treatment (baseline) and on day 28 as well as open-field activity on the 28 th day of the treatment. After the behavioral test, on day 28, mice were euthanized by cervical dislocation and the striatum were separated to determine tyrosine hydroxylase (TH), dopamine transporter (DAT) and glutamic acid decarboxylase (GAD) immunoreactivity by western blot analyses. Data are expressed as mean±standard error of the mean (SEM). Statistical analysis of data was carried out by one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test. Statistical significance was assumed at p<0.05 or less. The protocols were previously approved by the Ethics Commission on Animal Use of the Federal University of Santa Maria under process number 4562031214/2015. Results: The administration of gabapentin prevented the manifestation of haloperidol-induced VCMs. Treatment with haloperidol reduced locomotor activity in the open-field test which was reversed by gabapentin. We did not find any difference among the groups when tyrosine hydroxylase (TH), dopamine transporter (DAT) or glutamic acid decarboxylase (GAD) immunoreactivity parameters in the striatum were evaluated. Conclusion: These results suggest that the treatment with gabapentin, an analog of GABA, can attenuate the VCMs induced by haloperidol in mice.

Financial Support: CAPES, CNPq, FAPERGS.

Key words: tardive dyskinesia; locomotor activity; glutamic acid decarboxylase.

TAURINE MODULATES AGGRESSIVINESS INDUCED BY ETHANOL IN ZEBRAFISH ( Danio rerio ) Barbara D. Fontana

TAURINE MODULATES AGGRESSIVINESS INDUCED BY ETHANOL IN ZEBRAFISH (Danio rerio)

Barbara D. Fontana 1 , Daniele L. Meinerz 2 , Nathana J. Mezzomo 3 , Luiz Vinícius C. Rosa 1 , Vanessa A. Quadros 1 , Denis B. Rosemberg 1

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria. 2 Laboratório de Neuropsicobiologia Experimental, Universidade Federal de Santa Maria.

3 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria.

Introduction: Taurine (TAU) is an amino sulfonic acid with pleiotropic actions on brain function, which plays a role as neuromodulator, antioxidant, osmoregulator and membrane stabilizer. Ethanol (EtOH) is a drug that alters the functionality of GABA A receptor and modifies the balance of inhibitory/excitatory synapses, increasing agonistic behavior (e.g. aggressiveness) in animal models. Taurine is one of the most abundant molecules present in energy drinks frequently used as mixers for alcoholic beverages. However, the combined effects of TAU and EtOH on behavioral parameters such as aggression are poorly understood. Objective: The aim of this study was to investigate whether TAU modulates EtOH-induced aggression in zebrafish using the mirror-induced aggression (MIA) test. Methods: A total of 96 zebrafish (Danio rerio) were divided into 8 groups with X replicates [control (water); 42 mg TAU/L; TAU 150 mg/L; TAU 400 mg/L; 0.25% EtOH; TAU 42 mg/L in EtOH; TAU 150 mg/L in EtOH and TAU 400 mg/L in EtOH] to assess the effects of TAU pretreatment in the acute exposure of EtOH. After one hour of treatment, fish were placed individually in a tank with an inclined mirror for 6 minutes. Results: Fishes treated with EtOH, TAU 42, TAU 400, TAU 42/EtOH and TAU 400/EtOH showed increased aggression, while 150 mg/L TAU caused only an increased latency to attack the mirror. The 150mg/L TAU concentration also prevented EtOH-induced aggression, suggesting that it antagonizes the effects of acute alcohol exposure. Representative ethograms revealed the existence of different aggressive patterns and our results were confirmed by the index used to estimate aggression in the MIA test. Taurine alone did not alter pigment intensity, while EtOH and all co-treated groups presented a substantial increase in body color. Conclusion: These data suggest a biphasic effect of TAU on EtOH-induced aggression of zebrafish, which is not necessarily associated with changes in body color.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: aggressiveness; agonistic behavior; mirror-induced aggression test.

TRPA1 PARTICIPATION IN ANALGESIA TRIGGERED BY Tabernaemontana catharinensis ETHYL ACETATE FRACTION ON AN ARTHRITIC PAIN

TRPA1 PARTICIPATION IN ANALGESIA TRIGGERED BY Tabernaemontana catharinensis ETHYL ACETATE FRACTION ON AN ARTHRITIC PAIN MODEL IN MICE

Evelyne S. Brum 1 , Gabriela Beck 2 , Sara M. Oliveira 3

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

2 Laboratório de Neurotoxicidade e Psicofarmacologia, Universidade Federal de Santa Maria.

3 Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Santa Maria.

Introduction: The transient receptor potential ankyrin 1 (TRPA1) is an ion channels essential for detection and transmission of painful stimulus. Originally, TRPA1 was proposed as a sensor to noxious cold and has been identified as a potential target for the development of analgesics to treat painful inflammatory conditions such as rheumatoid arthritis induced by complete Freund's adjuvant (CFA). Tabernaemontana catharinensis (Apocynaceae) is a medicinal plant, popularly known as “cobrina”, used in the treatment of painful and inflammatory disorders. Several biological activities have been described for T. catharinensis ethyl acetate fraction (Eta) which are attributed to its polyphenol and flavonoid compounds. Objective: Considering that rheumatoid arthritis is a painful condition inadequately treated, we investigated the antinociceptive potential of T. catharinensis Eta on mechanical and cold allodynia in a rheumatoid arthritis model CFA-induced in mice. Methods: Adult male Swiss mice (30-35g) were used. The protocols were previously approved by the CEUA- UFSM (23076.038594/2014-57). The animals baseline (B1) paw withdrawal threshold (PWT) front the mechanical stimulus was verified using the “Up-and-Down” paradigm (Dixon, 1980). Next, the mice received 20 µL of CFA (1 mg/mL, i.pl.) and after 48 h, the baseline 2 PWT (B2) was verified. The animals were treated orally with Eta (100 mg/kg, v.o.) ou vehicle (10 mL/kg, v.o.). A significant decrease on B2 values vs. the B1 values were considered as mechanical allodynia. The mechanical threshold was evaluated from up 0.5 to 48 h after treatments. These animals also were utilized to evaluate the Eta or vehicle effect in front cold allodynia induced by an acetone drop (20 µL) applied three times into the dorsal surface of the right hind paw. An score (0-4 points) was determined in each application and the scores sum from up 0.5 to 48 h was used as final result. Results: The CFA treatment produced mechanical allodynia in mice treated with vehicle in all times evaluated. Eta (100 mg/kg) was effective in reversing the mechanical CFA-induced allodynia from up 0.5 to 24 h with maximal inhibition of 83.4±5.1% at 4 h after treatment. In addition, mice treated with Eta were capable of reduce (61.9±12.4%) the cold acetone-induced allodynia when compared with mice treated with vehicle. Conclusion: These results support the potential antinociceptive activity of T. catharinensis ethyl acetate fraction to treat the arthritic pain.

Financial Support: UFSM, CAPES.

Key words: cobrina; antinociception; inflammatory pain.

EPIDURAL KETAMINE, MORPHINE OR METHADONE FOR ACUTE PAIN IN BITCHES UNDER OVARIOHYSTERECTOMY Fernando S. F.

EPIDURAL KETAMINE, MORPHINE OR METHADONE FOR ACUTE PAIN IN BITCHES UNDER OVARIOHYSTERECTOMY

Fernando S. F. da Cruz 1 ; Fernanda Windmoller Brendler 2 ; Andréia Kusiack Gollo 2 ; Mauricio Borges da Rosa 3 ; Emanuelle Bortolotto Degregori 2

1 Docente do Curso de Medicina Veterinária, Departamento de Estudos Agrários, Universidade Regional do Noroeste do Estado do Rio Grande do Sul.

2 Aluno de Graduação em Medicina Veterinária, Universidade Regional do Noroeste do Estado do Rio Grande do Sul.

3 Médico Veterinário, Hospital Veterinário, Departamento de Estudos Agrários, Universidade Regional do Noroeste do Estado do Rio Grande do Sul.

Introduction: There is considerable evidence that pain associated with peripheral tissue or nerve injury involves NMDA receptor activation on primary afferent neurons. Ketamine, a dissociative anesthetic agent bind specifically to a site (PCP binding site) associated with the NMDA receptor monitored ion channel and inhibit the action of excitatory amino acid neurotransmitters at the NMDA receptor in a non-competitive manner. Morphine is the prototype opioid and is the agent of choice since is opioid agonist mu (µ) receptors exerting analgesic effect by reducing pain neurotransmitters. Methadone is a µ opioid agonist widely used to provide analgesia in small animals. It is a racemic mixture, although the analgesic action of the racemic mixture is primarily attributed to levomethadone, which is considered to have 10-50 times greater affinity for µ receptor than the dextrorotatory enantiomer. The latter is considered to have an NMDA receptor antagonist effect. Unlike morphine, methadone acts at different receptor systems, including variable affinity at NMDA and alpha 2 adrenergic receptors. Objective: The aim of this study was to compare the analgesic quality and cardiorespiratory effects of methadone, morphine or ketamine by epidural administration in bitches undergoing ovariohysterectomy. Methods: Eighteen bitches (16,1±3,9Kg) were randomly assigned to three groups, Methadone (GMe), Morphine (GMo) and Ketamine (GCe), receiving 0.2mg/kg; 0.1mg/kg and 2mg/kg respectively. After premedication with acepromazine 0.05mg/kg IM, Propofol 5mg/kg was given IV and intubation was performed to maintain anesthesia with isoflurane. When the animals were in adequate anesthetic plane, lumbosacral epidural was performed. All the volumes were adjusted to a volume of 0.75ml/kg with saline 0.9%. Heart Rate, Respiratory Rate, Systolic, Mean and Diastolic Arterial Pressure, SpO2 and o C was evaluated. Post operatory analgesia was evaluated by Visual Analogic Scale (VAS) and Glasgow Modified Scale (GMCS) at time 0, before any procedure and 0,5; 1; 1,5; 2; 3; 4; 5 and 6 hours after the end of the procedure. Results: The scores in post operatory evaluation by GMCS and VAS were significantly lower in GMe when compared with GMo and GCe. Bradycardia occur in two animals of GMe needing atropine administration, while RR reduced in all groups. Blood pressure, SpO2 and o C remain stable in all groups and GCe needed rescue analgesia during the post operatory evaluation. Conclusion: These data suggest that Methadone exerts a better quality of analgesia without great side effects when administered by epidural route in bitches submitted to ovariohysterectomy in the doses studied.

Financial Support: FAPERGS; PIBIC-UNIJUÍ

Key words: epidural; NMDA receptor; opioids.

EFFECTS OF AMPH-ISOTERAPIC ON AMPH DEPENDENCE PARAMETERS Hecson J. Segat 1 , Vinícia G. Metz

EFFECTS OF AMPH-ISOTERAPIC ON AMPH DEPENDENCE PARAMETERS

Hecson J. Segat 1 , Vinícia G. Metz 3 , Higor Z. Rosa 3 , Raquel C. S. Barcelos 2 , Marilise E. Burger 1,2,3

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria. 2 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria. 3 Centro de Ciências da Saúde, Universidade Federal de Santa Maria

Introduction: Addiction to psychostimulant drugs is a serious public health problem, involving dependence and relapses, which are related to deterioration of the subject, their family and society. Amphetamine (AMPH) and its derivatives are addictive drugs used due their capacity do promote alertness, motivation, willingness, courage and wellbeing. However, these drugs are related to negative consequences as insomnia, memory loss, paranoia, besides cardiac disorders. Currently, few effective therapies for the treatment of dependence on psychostimulant drugs are disponible, while the number of users grows alarmingly in worldwide. Objective:

Our aim was to evaluate the influence of the AMPH-isoterapic on AMPH-induced dependence parameters. This tool involves the addictive drug (AMPH) ultra-diluted and strengthened in proximate scale (10 -24 ). Methods: Male rats (Wistar, weighing, ±100g) were conditioned with amphetamine (AMPH) (4,0mg/Kg, i.p.) in conditioned place preference (CPP) paradigm for 8 days, resulting in two initial experimental groups (Saline and AMPH). Then, half of each experimental group was exposed to isoterapic treatment or placebo (alcohol 10ºGL) for the next 14 days, during AMPH extinction, resulting in four final groups (I-Saline/placebo; II-Saline/isoterapic; III-AMPH/placebo and IV-AMPH/isoterapic). At the sequence, animals received an additional dose of AMPH (2,0mg/kg, i.p.) and were submitted to the CPP-test in order to evaluate relapse behavior. Following behavioral assessments, prefrontal cortex, which is heavily involved in addiction, was be used for molecular analisys of dopamine transporter (DAT), tyrosine hydroxylase (TH) and dopamine receptors (D1 and D2) .Results: AMPH was able to induce preference in first CPP-test (without isoterapic treatment), however, 14 days of isoterapic treatment prevented AMPH relapse. Besides, AMPH increased DAT, TH, and D1 imunorreactivity, while isoterapic reduced these parameters. However the D2 imunoreactivity was decreased in both groups which received AMPH. Conclusion: At the moment, we recognize that in spite of therapeutic innovation, our studies need more data, and launch new alternative ideas on treatment for addition, also related to the use of AMPH-like drugs. Isoterapic treatment showed a promising tool on psychostimulant addiction, once, unfortunately, conventional pharmacological approach at the expense of tranquilizers and anxiolytics has low clinical efficacy, and high relapse rate, which favors the challenge of the proposals presented here.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: Addiction, AMPH, conditioning, isoterapic.

CHRONIC CONSUMPTION OF INTERESTERIFIED FAT MODIFIES BRAIN OPIOID SYSTEM AND AFFECTS MORPHINE-INDUCED REWARD EFFECTS IN

CHRONIC CONSUMPTION OF INTERESTERIFIED FAT MODIFIES BRAIN OPIOID SYSTEM AND AFFECTS MORPHINE-INDUCED REWARD EFFECTS IN RATS

Laura H. Milanesi 1 , Karine Roversi 1 , Caren D. Antoniazzi 1 , Hecson J. Segat 2 , Maikel Kronbauer 1 , Lívia F. D’Ávila 1 , Veronica T. Dias 1 , Marcel H.M. Sari 2 , Raquel C.S. Barcelos 1 , Fabíola Trevizol 1 , Marilise E. Burger 12 *

1 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria.

2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: Opioid analgesics are intensely used to alleviate acute and chronic pain, presenting high addictive potential and its improper use may be related to addiction, tolerance and withdrawal syndrome. Considering the high consumption of processed foods, interesterified fat (IF) has been used in replacement to trans fat since it causes deleterious health effects and may harm nervous system functions modifying addiction parameters. Objective: So far, no studies reported the influence of IF consumption on neuronal function, specially involving the opioid system and morphine, which became the aim of this study. Methods: Wistar rats were supplemented with soybean oil (SO) or IF during gestation, lactation and postweaning periods until pups’ adolescence. On postnatal day 39, animals were injected with morphine (4mg/Kg i.p.) in a conditioned place preference (CPP) paradigm. After the morphine-CPP test, anxiety-like symptoms, and thermal sensibility were evaluated, in elevated-plus maze (EPM) and hot plate test, respectively. Hippocampus and spinal cord were used for molecular markers assessments. The protocols were previously approved by the Ethics Commission on Animal Use of the Federal University of Santa Maria (nº041235/2016). Results SO group showed morphine preference during drug withdrawal while IF group showed no preference or withdrawal symptoms, presenting higher sensitivity than SO group to thermal stimuli. In addition, morphine conditioning increased dopamine D1 receptor and N-methyl-D-Aspartate receptor immunoreactivity in the hippocampus of SO group, while these molecular changes were not observed in IF group. Independently of morphine conditioning, IF group showed increased kappa opioid receptors immunoreactivity in the spinal cord, which also showed a negative correlation with thermal sensibility. Conclusion: We concluded that chronic consumption of foods rich in IF can affect neurotransmission resulting in loss of the rewarding effects induced by morphine administration in rats. This study contributes to a related warning to public health since it is showing the harmful influence of IF on dopaminergic and glutamatergic neurotransmission, which are confirmed by molecular markers, so affecting rewards system related to opioid activation.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: fatty acids; developmental periods; conditioned place preference.

MAJOR NEUROTRANSMITTER SYSTEMS IN DORSAL HIPPOCAMPUS AND BASOLATERAL AMYGDALA CONTROL SOCIAL RECOGNITION MEMORY L. Bühler

MAJOR NEUROTRANSMITTER SYSTEMS IN DORSAL HIPPOCAMPUS AND BASOLATERAL AMYGDALA CONTROL SOCIAL RECOGNITION MEMORY

L. Bühler 1 , C.G. Zinn 1 , N. Clairis 2 , L.E. Cavalcante 1 , C.R.G. Furini 1 , I. Izquierdo 1 , J.C. Myskiw 1

1 Memory Center, Brain Institute of Rio Grande do Sul, Pontifical Catholic University of Rio Grande do Sul (PUCRS) Porto Alegre RS, Brazil. 2 Département de Biologie, ENS Lyon Lyon France.

Introduction: Social recognition memory (SRM) is crucial to form social groups and for species survival. Objective: We examined the participation dopaminergic, noradrenergic and histaminergic systems into CA1 region of the dorsal hippocampus (CA1) and basolateral amygdala (BLA) on SRM consolidation. Methods:

Male Wistar rats (3 months old) were bilaterally implanted with guide cannulae in CA1 or BLA. 7 days after rehabilitation and 4 days of habituation (20 min session) in an open-field arena, animals underwent a training session of social discrimination paradigm, which consists in exposure to a juvenile co-specific (21 days old) for 1 hour. 24 hours after, animals were placed again in the same arena with familiar (F) juvenile and a second juvenile (novel, N) for 5 min (test session). Microinjections into CA1 (1.0 µL) or into BLA (0.5 µL) were carried out immediately after a training session. Protocols were previously approved by the Ethics Commission on Animal Use of PUCRS, process number: 13/00363. Results: Animals treated with protein synthesis inhibitor, anisomycin into either CA1, p<0.05 (time spent by Vehicle-treated animals exploring the new juvenile during test session, t Veh-N vs. time spent by Anisomycin-treated animals exploring the new juvenile during test session, t Ani-N), or BLA, p<0.001 (Veh-N vs. Ani-N), were not able to recognize familiar juvenile during retention test. When infused into CA1, β-adrenoceptor agonist, isoproterenol (10.0 g/side), p<0.05 (Veh-N vs. Iso-N), D1/D5 dopaminergic receptor antagonist, SCH23390 (1.50 g/side), p<0.01 (Veh-N vs. SCH-N) and H2 histaminergic receptor antagonist, ranitidine (17.5 g/side), p<0.05 (Veh-N vs. Rani-N in CA1), and when infused into BLA, β-adrenoceptor antagonist, timolol (1.0 g/side), p<0.05 (Veh-N vs. Tim- N), D1/D5 dopamine receptor agonist, SKF38393 (12.5 g/side), p<0.05 (Veh-N vs. SKF-N), and H2 histaminergic receptor antagonist, ranitidine (17.5 g/side) also hindered recognition of familiar juvenile 24 hours later. In all cases, the impairment was abolished by co-infusion of agonist plus antagonist. Conclusion:

The both CA1 and BLA play fundamental roles in the consolidation of SRM, but these roles are different in each structure depending on the involvement of receptors therein.

Financial Support: CNPq, CAPES.

Key words: social memory; basolateral amygdala; hippocampus.

GESTATIONAL STRESS DECREASE GLUCOCORTICOID RECEPTORLEVELS AND INCREASE OXIDATIVE PARAMETERS IN HIPPOCAMPUS OF ADOLESCENT

GESTATIONAL STRESS DECREASE GLUCOCORTICOID RECEPTORLEVELS AND INCREASE OXIDATIVE PARAMETERS IN HIPPOCAMPUS OF ADOLESCENT RATS

Luciana T. Vey 1 , Higor Z. Rosa 2 , Vinícia G. Metz 2 , Verônica T. Dias 3 , Lívia F. D’avila 3 , Marilise E. Burger 1,2,3

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria. 2 Departamento de Fisiologia e Farmacologia, UFSM-RS, Brasil 3 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria.

Introduction: A relationship between gestational stress (gest-S) and physiologic responses in rats has been shown in our previous studies, including hyper reactivity of the hypothalamic-pituitary-adrenal (HPA) axis, anxiety, and drug addiction. These effects were reflected on the reactive oxygen species (ROS) generation, which participates of normal physiological processes, but, when in excess, is related to oxidative damage to biomolecules and cellular structures. Objective: Here we investigated the influence of gest-S on biochemical and molecular parameters in rat’s hippocampus of adolescent offspring. Methods: Five rat dams were exposed to gest-S protocol for two weeks, while another group of dams (n=5) was not exposed to the stress protocol. On postnatal day (PND) 1, male pups were identified and two animals of each mother were assigned to gest- S group (n=10) or unhandled (UH) (n=10). Plasma corticosterone levels were quantified in both PND 9 and PND 50. Hippocampus was used for determination of lipid peroxidation (LP) and protein carbonyl (PC) levels, as well catalase (CAT) activity, an antioxidant defense. One part of the hippocampus was reserved for immunoreactivity of the glucocorticoid receptor (GR). Results: Gestational stress exposed animals showed increased LP and PC levels in hippocampus compared to UH. The activity of CAT did not change, which was similar to UH group, but GR levels decreased in hippocampus. Plasma corticosterone levels were increased in both pups and adolescent rats, in relation to UH group. Conclusion: Our findings show that gest-S exposure is able to cause deleterious influence on pups and this continue through adolescence, as evidenced by increased plasma corticosterone levels, decreased GR and increased oxidative damages in hippocampus, which is a brain area related to memory and cognition. Additional studies involving different types of memory together with anxiolytic drugs must be developed.

Financial Support: CNPq, CAPES and PRPGP.

IMPACT OF T102C 5-HT2A GENE POLYMORPHISM ON MORTALITY OF AMAZON RIPARIAN ELDERLY: A FIVE YEARS

IMPACT OF T102C 5-HT2A GENE POLYMORPHISM ON MORTALITY OF AMAZON RIPARIAN ELDERLY: A FIVE YEARS FOLLOW-UP STUDY

Lucas Maciel 1 , Fernanda Barbisan 2 , Ivana B. Manica da Cruz 2 , Tális de Oliveira Silva 2

1 Laboratório de Neurobiologia e Toxinologia, (LANETOX), Universidade Federal do Pampa. 2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: Since the average human life expectancy has increased, we can observe the impact of ageing and age-related disease on our society, especially in developing countries. Estimations pointed that by the year 2050 there will be about two million elderly people (sixty years or more). Today, in Brazil, it is estimated the existence of 17.6 million of seniors. The increase in the number of elderly people has increased the interest to understand the pathway of aging-related diseases. Studies report that the genes linked to aging are related to cellular processes such as cell survival, cell cycle regulation and others biochemical processes. Recent data show that pleiotropic genes are responsible to increase aging process. Serotonin (5-HT) is a pleiotropic molecule that exerts several functions on brain and peripheral tissues via different receptors. Among these receptors, 5-HT2A has an important role in body physiology. 5-HT2A gene presents some variations such as T102C single nucleotide polymorphism (SNP) that has been associated with risk of neuropsychiatric and vascular disorders. Objective: This study investigated the potential impact of 5-HT2A imbalance caused by genetic variations on human lifespan in Amazon riparian elderly. Methods: The study consisted in a prospective study in Amazon riparian elderly free-living in Maués city, AM-Brazil during a period of 5-years. Elderly (> 60 years old) of the Family Health Strategy Program (FHS-SUS) of Maués (AM) were invited to participate. Blood collection and samples were packed to genotyping for the T102 polymorphism 5-HT2A byPCR -RFLP. From 637 subjects included in July 2009 cohort, we genotyped 471 (male=209; female =262 (55.6%) with 72.3 ± 7.8 years old (60-100). Results: The T102C-SNP genotypes frequency was TT= 14.0% (n=66); CC= 28.0% (n=132) and CT= 58.0% (n=273); allelic frequency was C= 0.577 and T= 0.423. From 80 elderly that died in the period investigated we verified a higher number of dead TT-carriers (27.3%) and CC carriers (21.2%) when compared to heterozygous carriers (CT= 12.5%). Cox-regression analysis showed that association between T102C-SNP and elderly survival was independent of age, sex and other health variables. Conclusion: The results suggest that imbalance of 5-HT2A increases the mortality risk after 60 years old.

Financial Support: FAPERGS/CNPq

Key words: Serotonergic system; mortality risk; behavior.

EFFECT OF GUARANÁ CHEMICAL MATRIX COMPOUNDS ON THE REVERSAL OF ANTIPSYCHOTIC DRUG-INDUCED INFLAMMATION IN RAW

EFFECT OF GUARANÁ CHEMICAL MATRIX COMPOUNDS ON THE REVERSAL OF ANTIPSYCHOTIC DRUG-INDUCED INFLAMMATION IN RAW 264.7 LINEAGE MACROPHAGES

Moisés Henrique Mastella 1 , Fernanda Barbisan 2 , Verônica Farina Azzolin 2 , Pedro Antonio Schmidt Prado-Lima 3 , Gustavo Cardenas 4 , Euler Ribeiro 5 , Cibele Ferreira Teixeira 2 , Vitor Bueno 4 , Ivana Beatrice Mânica da Cruz 2 , Emilio Ivo da Cruz Jung 2 .

1 Curso de Ciências Biológicas, Universidade Federal de Santa Maria;

2 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria.

3 Instituto do Cérebro, Pontifícia Universidade Católica de Porto Alegre;

4 Curso de Medicina, Universidade Federal de Santa Maria;

5 Universidade Aberta da Terceira Idade, Universidade do Estado do Amazonas (UnATI/UEA).

Introduction: Antipsychotic drugs are used in the long-term treatment of psychiatric patients and, studies show that these drugs increase the risk of diseases such as obesity, dyslipidemia, diabetes mellitus type II and insulin resistance among other metabolic disorders. Some plants have an anti-inflammatory action such as guaraná (Paullinia cupana), fruit that contains as main compounds of its chemical matrix caffeine, catechin and theobromine. Objective: To evaluate in vitro the inflammatory effect of antipsychotics and the possible effect of reversion of inflammation by the compounds of the chemical matrix of guaraná. Methods: RAW 264.7 cells cultured under adequate and controlled conditions were treated with olanzapine, quetiapine, ziprazidone isolated and in association with the compounds of the guaraná chemical matrix: caffeine, catechin and theobromine and with the 3 compounds in association. After treatment the cells were incubated for 24 and 72 hours. The evaluation was carried out, thus evaluating the rate of cell proliferation through MTT (3- [4,5- dimethylthiazol-2-yl] -2,5-diphenyltetrazolic bromide) and the labeling with propidium iodide via the cell cycle. Results: All antipsychotics tested induced increased cell proliferation when compared to the control group. However, when the cells were activated with the antipsychotics and treated with the guaraná compounds, they showed a decrease in inflammation /proliferation at levels similar to the control group. Conclusion: The antipsychotics showed an increase in the cellular proliferation, indicative of inflammation, when the cells were exposed to the guaraná compounds, there was a reversion of this inflammatory effect. Further studies need to be performed to confirm our results, as this would be an initial step for further studies in order to make them a therapeutic option concomitant with traditional treatment.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: Guaraná; inflammation; antipsychotic.

EFFECT OF SUBCHRONIC TREATMENT OF CYCLOOXYGENASE-2 INHIBITORS ON PENTYLENETETRAZOL (PTZ)-INDUCED SEIZURES IN MICE Paula

EFFECT OF SUBCHRONIC TREATMENT OF CYCLOOXYGENASE-2 INHIBITORS ON PENTYLENETETRAZOL (PTZ)-INDUCED SEIZURES IN MICE

Paula Michelotti 1 , Fernanda Rossatto Temp 1 , Joseane Righes Marafiga 1 , Ana Cláudia Jesse 1 , Laura Hautrive Milanesi 1 and Carlos Fernando Mello 1

1 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria.

Introduction: Cyclooxygenase-2 (COX-2) inhibitors reduce prostaglandins synthesis, and play a significant role in inflammation and fever. In the past decades, research has been focused on the use of COX-2 inhibitors and the inflammatory process in the central nervous system. However, there is a dispute regarding the role of COX-2 in acute and chronic seizures. Some studies have shown that COX-2 inhibition decreases seizures, while others have reported that it may facilitate convulsive episodes. Objective: Thus, considering the current discrepancy regarding the pro- and anticonvulsant effect of COX-2 inhibitors, and the use of different drugs, the aim of the current study was to investigate whether chronic administration of nimesulide, celecoxib and etoricoxib alter seizures in mice. Methods: Adult male Swiss mice received vehicle (0.1% carboxymethylcellulose plus 5% Tween 80, p.o.), nimesulide, celecoxib or etoricoxib (0.2, 2 or 20 mg/kg, p.o.), daily for 14 successive days. On the 15th day mice were challenged with PTZ (50 mg/kg, i.p.). After PTZ administration animals were monitored for 20 minutes for the appearance of myoclonic jerks, generalized tonic-clonic seizures, number of seizure episodes, total time spent seizing and Racine scale score. The protocols were previously approved by the Ethics Commission on Animal Use of the Federal University of Santa Maria under process number 024/2014. Results: The administration of nimesulide and etoricoxib significantly increased the latency to PTZ-induced generalized tonic-clonic seizures. However, celecoxib significantly decreased the latency to PTZ-induced generalized tonic-clonic seizures. Furthermore, nimesulide significantly decreased total time spent seizing, but did not alter the latency to PTZ-induced myoclonic jerks, number of seizure episodes and Racine scale score. Nevertheless, etoricoxib and celecoxib did not alter the latency to PTZ-induced myoclonic jerks, number of seizure episodes, total time spent seizing and Racine scale score. Conclusion: Our results suggest that the effect of COX-2 inhibitors on seizures may vary depending on the drug and dose administration. Such a discrepancy between the effects of COX-2 inhibitors may be due to differential COX-2 selectivity, putative interaction with other targets or pharmacokinetic issues.

Financial Support: CAPES, CNPq, FAPERGS, PRPGP/UFSM, PIBIC/UFSM.

Key words: seizure, nimesulide; celecoxib; etoricoxib; neuroinflammation.

MODAFINIL DURING ADOLESCENCE MODIFIES AMPHETAMINE- CONDITIONED PLACE PREFERENCE IN YOUNG ADULT RATS Verônica Tironi Dias

MODAFINIL DURING ADOLESCENCE MODIFIES AMPHETAMINE- CONDITIONED PLACE PREFERENCE IN YOUNG ADULT RATS

Verônica Tironi Dias 1 , Higor Zuqueto Rosa 3 , Lívia Ferraz D’avila 1 , Luciana Taschetto Vey 2 , Raquel Cristine Silva Barcelos 1 , Marilise Escobar Burger 1,2,3

1 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria.

2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

3 Departamento de Fisiologia e Farmacologia, Universidade Federal de Santa Maria.

Introduction: Addiction is a serious health problem leading to social impairments in general. The adolescent period of development plays a significant role in drug abuse liability. In view of that, the misuse of psychostimulants drugs, such as modafinil (MOD), mainly because of common use between healthy subjects that want prolonged wake state and improvements in cognition contribute to the indiscriminate use. Along with that, there is still limited knowledge about MOD neural mechanism of action. Objective: The present study aims to investigate the effects of MOD administration in adolescent rats on conditioned place preference (CPP) to amphetamine (AMPH) in young adulthood. Also, we evaluate the oxidative status and the D1R immunoreactivity of hippocampus. Methods: 28 Wistar male rats (PND21) were acclimated and received MOD (64mg/kg p.o., once a day) during 14 days (PND 28-42), these animals were not perturbed until young adulthood. At PND 60 they were submitted to AMPH-CPP protocol lasting 8 days. After behavioral evaluation, the animals were euthanized (under anesthesia) and the hippocampus was dissected for analysis. Results:

MOD interestingly decreased AMPH-CPP. MOD per se decreased reactive species (RS) generation and after AMPH exposure RS levels were lower than AMPH group. Also, MOD restored catalase (CAT) activity AMPH impaired. Besides that MOD per se has no influences on D1R levels and after AMPH exposure decreased D1R levels. Conclusion: So far, our initial hypothesis was not confirmed. The animals that received MOD during adolescence showed reduced AMPH-CPP in young adulthood. Also, MOD improved oxidative status, thus it was able to modulate D1R levels in hippocampus. These unexpected MOD behavioral effects on CPP could be related to the stable hippocampal oxidative status without alterations on D1R immunoreactivity, although, further studies are necessary.

Financial Support: CAPES, PROAP/ PRPGP (UFSM).

Key words: addiction; oxidative stress; dopamine receptor D1.

FISH OIL SUPPLEMENTATION REDUCES AMPHETAMINE-INDUCED OXIDATIVE DAMAGES Vinícia G. Metz 2 , Hecson J. Segat

FISH OIL SUPPLEMENTATION REDUCES AMPHETAMINE-INDUCED OXIDATIVE DAMAGES

Vinícia G. Metz 2 , Hecson J. Segat 1 , Marilise Escobar Bürger 1

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria. 2 Departamento de Fisiologia e Farmacologia, Universidade Federal de Santa Maria- UFSM-RS, Brasil.

Introduction: Drug dependence is a serious problem of public health, whose treatment options are scarce. Amphetamine-like drugs are psychostimulants clinically used, however, their recreational use have increased alarmingly, causing concerns due to their association with reinforcement, tolerance and dependence. These addiction parameters are also related to frequent relapses, besides cellular oxidative damage in brain areas. The chronic consumption of omega-3 polyunsaturated fatty acids (PUFA-3) had exerted beneficial influences against central nervous system disturbances, including drug addiction. Objective: Our aim was to evaluate influences of fish oil (FO, rich in PUFA-3) supplementation on AMPH-induced oxidative parameters. Methods: Wistar rats received d,l-AMPH (4 mg/kg, i.p) or saline for 14 days. Then, one half of each experimental group (AMPH or saline) was treated with FO (3 g/kg/day) or vehicle for fourteen days, when animals were re-exposed to AMPH or vehicle for more three days. After 24h of AMPH re-exposure, rats were euthanized and ventral tegmental area (VTA) was removed for biochemical analysis [Catalase (CAT) activity, Protein carbonyl (PC) levels and Reactive species (RS) generation]. Results: AMPH per se increased CAT activity, PC levels and RS generation in VTA while FO treatment was able to reverse these parameters. Conclusion: We observed that FO supplementation decreased AMPH-induced oxidative damages in the VTA. This supplementation may be considered a adjuvant therapeutic to treat addiction to psychostimulant drugs.

Financial Support: CNPq, FAPERGS.

Key words: Psychostimulants; Omega-3; Oxidative stress.

FINAL PROGRAM ABSTRACTS SESSION: BEHAVIORAL BIOLOGY

FINAL PROGRAM

ABSTRACTS

FINAL PROGRAM ABSTRACTS SESSION: BEHAVIORAL BIOLOGY

SESSION: BEHAVIORAL BIOLOGY

MODULATION OF GROOMING BEHAVIOR INDUCED BY THE UREASE OF C anavalia ensiformis (JBU) IN N

MODULATION OF GROOMING BEHAVIOR INDUCED BY THE UREASE OF Canavalia ensiformis (JBU) IN Nauphoeta cinerea COCROACHES

Ana Paula Zanatta 1 , Douglas Silva dos Santos 1,2 , Carlos Gabriel Moreira de Almeida², Maria Eduarda Rosa 1 , Bruna Trindade Borges 1 , Yuri Correia Barreto 1 , Lucas Maciel 1 , Allan Pinto Leal 1 , Barbara Akemi Ogata 1 , Etiely Karnop 1 , Lúcia do Canto Vinadé 1 , Célia Regina Carlini 3 , Cháriston André Dal Belo 1,2,3

1 Laboratório de Neurobiologia e Toxininologia (LANETOX), Universidade Federal do Pamp 2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

3 LANEUROTOX, Instituto do Cérebro, Pontifícia Universidade Católica do Rio Grande do Sul

Introduction: Ureases are metalloenzymes witch hydrolyze urea to ammonia and carbon dioxide.These compounds are synthesized by many organisms and have been characterized as natural insecticides. The entomotoxic activity of the urease of Canavalia ensiformis (Jack Bean Urease, JBU) has been proved, although the exact mechanism of action is still a matter of debate. Objective: The aim of this work was to investigate the mechanisms involved in the entomotoxic activity of JBU over the central nervous system of Nauphoeta cinerea cockroaches, using the grooming behavior analysis. Methods: N. cinerea (Olivier) of both sexes, were reared at 23-26°C, with water and dog chow ad libitum. Grooming activity was measured and analyzed as described elsewhere (Sturmer et al. 2014, Ecotoxicol Environ Saf. 109:56-62). Data were expressed as the mean ± S.E.M of the total number of leg and antenna groomings in seconds/30min. Treatments (10µl) were administered at the third abdominal segment of the cockroach, with a Hamilton syringe. Significances were taken at p<0.05, by the Student "t" test. In control parameters the values of leg grooming was 150 ± 5s/ 30min and for antennae 70 ± 8s/ 30min (n = 30), respectively. Results: When JBU (1.5, 3 and 6 µg/g of animal) was administered, there was a dose-dependent alteration in the grooming activity. Thus, at 1.5 µg/g of animal weight JBU, no alteration in the grooming activity was observed (n = 30). When 3 µg/g JBU was assayed, there was an increase in the grooming activity to 253 ± 30s for the legs (p<0.05), without any significative change in the antennae behavior (57± 7s/30min), n=30 respectively. At 6 µg/g JBU, there was a further increase in the leg grooming activity to 364 ± 23s/30min (p<0.05), without any modulation of the antennae counterpart (80 ± 9s/30min), n=30 respectively. The previous treatment of the animals with phentolamine (5uM), a selective octopamine receptor blocker, prevented significantly the increase of the leg grooming activity (6 µg/g JBU, 55± 9s/30min, n=28, p<0.05). Conclusion: The entomotoxic activity of Jack Bean Urease involves a modulation of the insect grooming behavior. The increase of leg grooming activity, together with the inhibition of this activity by phentolanine, suggests an octopamine-related mechanism by the urease entomotoxic activity.

Financial Support: Edital Toxinologia 063/2010 CAPES.

Key words: Natural Insecticides; Jack Bean Urease; Insect Behavior.

EFFECT OF DIETARY SUPPLEMENTATION OF TIGER NUT (C yperus esculentus L.) AND WALNUT Tetracarpidium conophorum

EFFECT OF DIETARY SUPPLEMENTATION OF TIGER NUT (Cyperus esculentus L.) AND WALNUT Tetracarpidium conophorum MÜLL. ARG.) ON SEXUAL BEHAVIOR, HORMONAL LEVEL AND ANTIOXIDANT STATUS IN MALE RATS

Ayodeji Augustine Oabiyi 1, 2, 3 , Ganiyu Oboh 1 , Stephen Adeniyi Adefegha 1

1 Federal University of Technology, Akure. Ondo State. Nigeria.

2 Afe Babalola University, Ado Ekiti. Ekiti State. Nigeria.

3 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: Tiger nut and walnut have been used in folklore medicine in the management/prevention of erectile dysfunction (ED) without scientific basis. Also, some pathological states have been linked to ED which includes hypertension, diabetes and other degenerative diseases. These nuts can be eaten either as raw or in processed form (roasted or cooked). Objective: The aim of this study is to assess the effect of dietary supplementation of tiger nut (Cyperus esculentus L.) and walnut (Tetracarpidium conophorum Müll. Arg.) on sexual behavior, hormonal level and antioxidant status in normal adult male rats. Methods: The animals (male and female) were divided into five groups of eight animals (n = 8). The male rats were grouped as follows:

Group 1 normal control rats, fed with basal diet; Group 2 rats fed basal diet plus 10% processed tiger nut; Group 3 rats fed diet supplemented with 10% raw tiger nut; Group 4 rats fed diet supplemented with 10% processed walnut; Group 5 - rats fed diet supplemented with 10% raw walnut. Daily feed intake was monitored. The experiment lasted 14 days after which the male and female animals were subjected to sexual (male paired with female) and anxiety (male) behavioral studies. Sexual behaviors were monitored in a separate room for 1h in a clear plastic box (60 × 60 × 80 cm) and elevated plus maze (anxiety) which was recorded by digital video recording and analysed. The male rats were euthanized with isoflurane, their blood collected for serum preparation followed by determination of sex hormone levels, enzymatic and non-enzymatic activities. The protocols were previously approved by the Ethical Committee on Animal Use of the Federal University of Technology, Akure. Results: The groups with supplemented diet containing tiger nut and walnut showed significantly increased sexual behavior, hormone levels and antioxidant activities. Conclusion: The results suggest the potential ability of tiger nut (Cyperus esculentus L.) and walnut (Tetracarpidium conophorum Müll. Arg.) to promote erectile function and be useful functional foods for both animal and human nutrition.

Financial Support: None.

Key words: functional food; sexual behavior; erectile function.

NEW MOLECULE DERIVED FROM 1, 3, 4-OXADIAZOLE MODULATES GABAERGIC SYSTEM AND PROTECTS AGAINST CONVULSION-LIKE BEHAVIOR

NEW MOLECULE DERIVED FROM 1, 3, 4-OXADIAZOLE MODULATES GABAERGIC SYSTEM AND PROTECTS AGAINST CONVULSION-LIKE BEHAVIOR IN Caenorhabditis elegans

Daniela Freitas Câmara¹, Marina Lopes Machado¹, Letícia Priscilla Arantes¹, Thayanara Cruz da Silva¹, Tássia Limana da Silveira¹, Fabiane Bicca Obetine Baptista¹, Alisson Vinholes¹, Félix Alexandre Antunes Soares¹.

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: Seizures can be described as paroxysmal hypersynchronous transient electrical discharges that result from too much excitation or too little inhibition in the area in which the abnormal discharge starts. Currently 50 million people have seizures and one third of these patients are refractory to available treatments due to their side effects, requiring the search for new drugs. Derivatives of 1,3,4-oxadiazoles compounds have been reported as positive modulators of the GABAergic system, and showed anticonvulsant activity in Pentylenetetrazole (PTZ)-induced seizure models. Objective: Evaluate the protective effects of 2-((((4- methoxyphenyl) selenil) methyl) thio) -5 phenyl-1, 3, 4-oxadiazole against PTZ-induced seizures in the nematode Caenorhabditis elegans. Materials and methods: Transgenic worms with a reduction in GABAergic signaling (unc-25, unc-30, unc-46, unc-49 e unc-47) were exposed to DMSO vehicle (control) or

15 µM of the oxadiazole compound, from the larval stage (L1) to young adults. After 48 hours of treatment,

we measured the latency time of the worms to the onset of seizures and the time to recovery after induction of

seizures with 5mg / ml of agar of PTZ. Results: The worms with deletions in the unc-30, unc-46 and unc-49 gene treated with 15 µM of the compound showed an increase of approximately 50% in the latency time until the occurrence of seizure. They also moved more after seizures, showing an increase of approximately 20% in the number of body bends per minute compared to the control group. Worms unc-25 and unc-47 showed no response to treatment with the compound. Conclusion: The compound was effective in the protection and recovery of worms with deletions in the genes unc-30 (acts on axonal growth and control of synapses), unc-

46 (regulate the transport of GABA into vesicles) and unc-49 (encodes the GABAA receptor) on seizures

induced by PTZ. However, it was not active in the absence of the gene that regulates the synthesis of GABA (unc-25) and the gene encoding this neurotransmitter vesicle carrier (unc-47). Therefore, our data suggest that the oxadiazole compound tested may modulate inhibitory system (GABAergic) in an unc-25 and unc-47- dependent manner. More studies are needed in order to better understand the mechanisms of action of this molecule.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: Caenorhabditis elegans; convulsion; GABAergic system.

EVALUATION OF LACTATE DEHYDROGENASE ACTIVITY IN ADULT ZEBRAFISH UNDER PRECONDITIONING CONDITIONS Emerson Santos da Silva

EVALUATION OF LACTATE DEHYDROGENASE ACTIVITY IN ADULT ZEBRAFISH UNDER PRECONDITIONING CONDITIONS

Emerson Santos da Silva 1 , Marcos Martins Braga 2 , Maria Elisa Calcagnotto 2

1 Biochemistry Department, NNNESP Lab, UFRGS, Porto Alegre, RS, Brazil; 2 Programa de Pós-Graduação em Bioquímica Toxicológica, Departamento de Bioquímica e Biologia Molecular, Centro de Ciências Naturais e Exatas, Universidade Federal de Santa Maria, Santa Maria, RS, Brazil.

Introduction: One of the most common phenomena in brain diseases is the lack of brain oxygenation.

Thus, our group began the study on new therapeutic approach in hypoxia-related diseases. The preconditioning (PC) under low O 2 levels has been studied as therapy for the treatment of hypoxia. Objectives: Thus, we measured the activity of lactate dehydrogenase (LDH) in different tissues (brain and blood) post-PC in order

to study the effect of this treatment to understand the mechanisms of hypoxia tolerance. Methods: To induce the PC, adult zebrafish were placed in a glass chamber to reduce the O 2 levels by addition of nitrogen gas. The

O

2 was kept at a low level (1.8 mg/L for 10 minutes) in the severe PC group and between 3.5-4.0 mg/L for 2

h

in the moderate PC group. After PC the animals were separated into different groups for recovery under

normoxia (~8.0 mg/L) at 6, 12, 24, 48, and 72h. Next, the animals were anesthetized and euthanized for brain

dissection and blood collection, to evaluate the LDH activity. Results: LDH activity in the brain was increase

in animals exposed to severe PC at 6, 12 and 24h. There was no difference in LDH activity in the blood between

groups. The animals exposed to moderate PC showed an increase in the brain LDH activity after 12 and 24h and in the blood after 12, 24, 48 and 72h. Conclusions: The increased brain LDH activity may be related to the high brain energetic demand. The increased LDH activity in the blood in animals exposed to moderate PC could be attributed to the longer period of PC (2h rather than 10min). Therefore different mechanisms of tolerance to hypoxia could be involved in the two distinct PC conditions that should be further explored.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: lactate dehydrogenase; preconditioning.

BEHAVIORAL EVALUATION AFTER ACUTE AFLATOXIN B1 EXPOSURE IN RATS Érica Furlan da Rosa 1 ,

BEHAVIORAL EVALUATION AFTER ACUTE AFLATOXIN B1 EXPOSURE IN RATS

Érica Furlan da Rosa 1 , Naiéli Schiefelbein Souto 2 , Micheli Dassi 2 , Ana Claudia Monteiro Braga 1 , Ana Flávia Furian 1,2

1 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria 2 Programa de Pós-Graduação em Ciência e Tecnologia em Alimentos, Universidade Federal de Santa Maria.

Introduction: Aflatoxins (AF) represent an important group of harmful mycotoxins commonly found as contaminants in grains and peanuts. Food contamination by AF continues to be a significant public health problem worldwide, particularly in developing countries. Aflatoxin B1 (AFB1) is not only the most toxic but also the most abundant of aflatoxins. Many researches clarified the peripheral effects of the exposition to AFB1, however, there are few studies explaining their effects on central nervous system. Objective: evaluate the acute effects on behavior caused by the oral administration of AFB1. Methods: Young male Wistar rats received a single administration of AFB1 (250 µg/kg/i.g.) and 48h after they were submitted to behavioral analysis: open-field test, novel object recognition, forced swim test, taste preference test, elevated plus maze. The protocols were previously approved by the Ethics Commission on Animal Use of the Federal University of Santa Maria under process number 093/2014. Results: Mycotoxin in this protocol was not able to modify the behavior of the animals when compared to the control group, and the primary damage is probably signaled by biochemical and non-behavioral mechanisms. Conclusion: In this acute protocol AFB1 did not induce any behavioral change.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: mycotoxin; behavioral analysis; Food contamination; rat.

THE EVOLUTIONARY HISTORY OF A VIRAL GLYCOSYLTRANSFERASE THAT CHANGES HOST BEHAVIOR DURING VIRUS INFECTION Ethiane

THE EVOLUTIONARY HISTORY OF A VIRAL GLYCOSYLTRANSFERASE THAT CHANGES HOST BEHAVIOR DURING VIRUS INFECTION

Ethiane R. Santos 1,2 , Lucas B. Oliveira 2 , Lenen Peterson 2 , Daniel Ardisson-Araújo 1,2

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria. 2 Laboratório de Virologia de Insetos, Universidade Federal de Santa Maria.

Introduction: Viruses are cell-obligate toxic agents that can alter the host behavior. For instance, caterpillars infected by several baculviruses migrate to the upper foliage of the host plant where they die and liquefy. This climbing up symptom is associated with the expression of a glucosyltransferase (egt) gene whose product inactivates molting and keeps the insect into the larval stage. Baculoviruses are divided into four genera, two of them Alphabaculovirus and Betabaculovirus (subdivided into clade-A and B) are infectious to moths and butterflies. Importantly, the evolutionary history of the egt in these groups is not clear. Objective: To identify egt genes in five novel baculovirus genomes (SpcoNPV, DijuMNPV, MyseNPV, UrprNPV, and MospGV) and reconstruct the evolutionary history of the gene to understand the insect climbing up behavior. Methods: we sequenced the genome of five new viruses using 454-pyrosequencing. The genomes were assembled and annotated using Geneious-R9 and BLAST-X. We also reconstructed the virus phylogeny using a set of conserved genes and prospected for the presence and absence of the egt in all baculovirus sequenced. The egt sequences found were aligned based on a PDB-crystal structure to perform a phylogenetic reconstruction. Results: we found that egt is a viral gene acquired from an insect host. All alphabaculoviruses presented egt whereas betabaculovirus varied, some presented and some did not. Regarding the new genomes, solely UrprNPV presented an egt with high genetic diversity in comparison to the other viruses and MospGV lacked a homolog. Remarkably, we also found that alphabaculovirus possibly transferred the gene to a betabaculovirus ancestor then the virus lineages underwent three independent losses: an ancestral loss in clade- A (that contains MospGV) and two independent losses in viruses from clade-B i.e. DisaGV and CnmeGV. Conclusion: The influence of egt on the climbing up behavior is unclear and its evolutionary history in betabaculovirus has never been shown. The egt might have been product of a horizontal transfer from alphabaculovirus during a co-infection scenario followed by three independent losses. Interestingly, in the case of DisaGV and CnmeGV lacking gene both of them infect caterpillars that have tunnels-living habits inside the plant and do not present the climbing up symptom.

Financial Support: CNPq, FAPDF.

Key words: baculovirus; glycosyltransferase; tree-top disease.

Nauphoeta cinerea AS ALTERNATIVE MODEL FOR STUDY THE EFFECT OF PRALIDOXIME ON BEHAVIORAL IMPAIRMENT INDUCED

Nauphoeta cinerea AS ALTERNATIVE MODEL FOR STUDY THE EFFECT OF PRALIDOXIME ON BEHAVIORAL IMPAIRMENT INDUCED BY CHLORPYRIFOS

Fernanda D’Avila da Silva 1 , Marcos Martins Braga 1 , João Batista Teixeira da Rocha 1,2

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria. 2 Departamento de Bioquímica e Biologia Molecular, CCNE, Universidade Federal de Santa Maria.

Introduction: Indiscriminate use of organophosphate (OP) pesticides generates risks associated with occupational exposure. OPs are potent and irreversible inhibitors of acetylcholinesterase (AChE), which is the enzyme responsible by the hydrolysis of the neurotransmitter acetylcholine (ACh). AChE inhibition causes the accumulation of ACh in synaptic cleft, leading to overstimulation of cholinergic receptors associated to locomotor depression, paralysis, convulsion and eventual death. In mammals, the treatment for intoxication by OPs includes the administration of atropine, oximes and anticonvulsive agents. However, there is limited information describing the acute neurobehavioral effects of chlorpyrifos (CPF) and its reversion by oximes using invertebrate models. The AChE isoforms from insects present similarities with those from vertebrates and, consequently, insects could be exploited as an alternative model for studying the interaction of CPF and oximes on AChE activity. Objective: This study has the aim of sought a new alternative model for research, and analysis behavioral of cockroaches treated with pralidoxime (P) and CPF. Methods: Male cockroaches were separate in four groups (N=10 per group): Control, CPF, P and CPF followed by P (CPF + P). First, the animals from the Control and P groups were exposed to saline (10 µL/cockroach) and the animals from the CPF and CPF + P groups were exposed to CPF 3.4 nmol/10 µL/cockroach. After 10 minutes, the Control and CPF groups were treated with saline (10 µL/cockroach), and P and CPF + P groups were treated with pralidoxime 120 nmol/10 µL/cockroach. The compounds and saline were administrated via an injection in the body. After, the cockroaches were immediately transferred to a white cage (10 cm in width × 10 cm in length × 1.5 cm in height) in order to evaluate their spontaneous behavior during 60 min. The behavioral profiles were determined by the main behavioral phenotypes such as normal behavior, erratic movements, seizure-like behavior and loss of posture. Results: The CPF injection significantly impaired the behavioral repertory of cockroaches and the administration of pralidoxime 10 min after CPF reversed almost completely the behavioral changes induced by CPF. Conclusion: Thus, it became clear that: 1) cockroaches are sensitive to CPF; 2) pralidoxime was able to reverse the toxic effects caused by CPF; and 3) cockroaches can be a potential experimental model for research.

Financial Support: CAPES, CNPq, FAPERGS.

Key words: cockroaches; behavior; organophosphate; oxime.

PHYSICAL EXERCISE AND BRAIN REPERCUSSION Fernandes, Carlos A. O. R.¹,(IC); Oliveira, Renata S. C.²(IC); Sangoi,

PHYSICAL EXERCISE AND BRAIN REPERCUSSION

Fernandes, Carlos A. O. R.¹,(IC); Oliveira, Renata S. C.²(IC); Sangoi, Larissa G.²(IC); Bordignon, Eduardo M.² (IC); Daronco, Luciane S.E.¹ (O)

1 Departamento de Educação Física Bacharelado, Universidade Federal de Santa Maria. 2 Departamento de Ciências da Saúde, Medicina, Universidade Federal de Santa Maria.

Introduction: For decades, a beneficial relationship between physical exercise and central nervous system metabolism has been demonstrated, after a single session of aerobic exercise performed at moderate intensity, improvements in the performance of different cognitive tasks such as processing speed, selective attention and short-term memory are observated. It has recently been demonstrated that the brain is responsive to physical activity, that is, it has potential in the prevention and treatment of traumatic brain damage, as well as in neurodegenerative diseases such as Parkinson's disease (PD) and Alzheimer's disease, in addition, studies support that many of these changes occur in specific areas of important brain functions such as long-term memory and prevention of cognitive decline during aging. The regularity of the practice of physical activities, for example, benefits individuals with Parkinson's disease, having an important role to soften or delay the onset of symptoms, guaranteeing some independence for those affected; Mainly aerobic physical exercise is beneficial for PD patients, because it reduces symptoms such as bradykinesia, hypokinesia and gait disorders, being an important tool in the aid of drug therapy. Physical exercise may interfere with cognitive performance for a number of reasons, such as increased levels of neurotransmitters and changes in brain structures, cognitive improvement observed in individuals with mental impairment, and limited improvement in elderly individuals, due to lower mental and attentional flexibility. In addition, chronic stress can lead to constant high levels of cortisol in the blood, which in addition to interfering with increased appetite and depression of the immune system, can cause sleep disturbances, loss of libido, elevate heart rate, blood pressure and levels of fat in the blood. Objective: To describe the cerebral repercussion and the benefits of physical exercise in the cognition of people with and without neurodegenerative diseases. Methods: A bibliographic review was done from the year 2006 to 2014 in order to approach characteristics described by several authors on a subject of great importance. Conclusion: Individuals with and without neurodegenerative diseases should perform a daily routine of physical exercise to avoid a series of negative repercussions in the brain, as well as help in the prevention of numerous diseases.

Financial Support: None.

SODIUM SELENITE IS A NEUROPROTECTIVE AGENT AGAINST BEHAVIORAL CHANGES INDUCED BY PARAQUAT IN ZEBRAFISH Flavia

SODIUM SELENITE IS A NEUROPROTECTIVE AGENT AGAINST BEHAVIORAL CHANGES INDUCED BY PARAQUAT IN ZEBRAFISH

Flavia Vestena Stefanello 1 , Talise Ellwanger Müller 1,2 , Mauro Medina Nunes 2 , Denis Broock Rosemberg 1,2 , Vania Lucia Loro 2

1 Laboratório de Neuropsicobiologia Experimental, Universidade Federal de Santa Maria.

2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: Brazil is one of the largest consumers of herbicides in the world, including paraquat (dichloride 1,1'-dimethyl-4,4'-bipyridine, PQ). PQ can enter the central nervous system causing a selective destruction of dopaminergic neurons, changing the operation of enzymes involved in dopamine metabolism, which may be directly associated with the development of neurodegenerative diseases such as Parkinson's disease. Therefore, a selenium (SE) supplementation can be used as a strategy to prevent the oxidative damage caused by PQ. SE has antioxidant and neuroprotective activities playing an important role in the proper functioning of cells because it is an essential micronutrient for maintaining normal brain function in vertebrates. SE supplementation can prevent motor and non-motor disorders as cognitive decline, mood disorders, and

functional deficits in several neurodegenerative diseases. Objective: To investigate whether Na 2 SeO 3 dietary prevents behavioral changes triggered by PQ in zebrafish. Methods: In this experiment, the fish were

Na 2 SeO 3 for 21 days. Fish were further subdivided in four

pretreated with a diet supplemented with 1 mg/kg

experimental groups: control group (CT), paraquat group (PQ), Na 2 SeO 3 group (SE) and selenium + paraquat group (SE+PQ). PQ (20 mg/kg) was administered intraperitoneally, totalizing 6 injections for 16 days. The behavioral tests were performed 24 hour after the last injection. Results: In the novel tank test, Na 2 SeO 3 diet prevented the locomotor impairments, as well as the increased the time spent in top area of tank, and the exacerbation of freezing episodes. In the preference for conspecifics and in the mirror-induced-aggression

(MIA) tasks, Na 2 SeO 3 prevented the increase in the latency to enter the area closer to conspecifics and the agonistic behavior of PQ-treated animals, respectively. Conclusion: The Na 2 SeO 3 dietary prevented behavioral changes triggered by PQ in zebrafish because it modulates anxiety- and aggressive-like phenotypes in zebrafish.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: Selenium; Zebrafish; Behavioral; Parkinson’s disease.

EFFECT OF RESERPINE ON THE AVOIDANCE OF NOXIOUS HEAT BEHAVIOR IN Drosophila melanogaster Getulio N.

EFFECT OF RESERPINE ON THE AVOIDANCE OF NOXIOUS HEAT BEHAVIOR IN Drosophila melanogaster

Getulio N. Bressan 1 , Roselei Fachinetto 1,2

1 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

2 Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Maria.

Introduction: The Drosophila melanogaster, known as fruit fly, is used in scientific research as a model organism because it has advantages over mammals. The nociception is warning mechanism to organism when this is facing a harmful agent in the environment avoiding damage to your body. The dopamine is a catecholaminergic neurotransmitter is involved in several physiological processes in flies, such as locomotion, sensitivity and thermal body preference. Reserpine is a vesicular monoamine transporter inhibitor and had been used to reduce endogenous dopamine levels in D. melanogaster. Objective: The aim of this study was to investigate if reserpine influences on the avoidance of noxious heat behavior (ANH) in D. melanogaster. Methods: First, it was determined the temperature which is noxious to the fly by thermal tolerance profile (TTP). D. melanogaster were treated with reserpine (3, 10, and 30 μM) or control (acid acetic 1%) in the treatment medium of agar during three days. After 72h, flies were separated to experiments of negative geotaxis (10 flies) and ANH behaviors (20 flies). In the negative geotaxis test was verified the number of flies crossing 6 cm at 6s, and ANH behavior was evaluated the number of flies paralyzed on basis of Petri dish after 4 min at 46ºC. Results: Through TTP was verified that the 46ºC temperature was harmful to the fly. Flies treated with 10 and 30 μM of reserpine showed impairment of locomotion activity. However, only the concentration of 30 μM significantly decreased the ability to escape the noxious heat and thus maintaining the flies on the basis of the Petri dish. This effect of reserpine on the ANH behavior seems to have no interference on decreasing locomotor activity of the fly since the flies treated with 10 μM showed loss of locomotion but without a decrease in the ability to escape the noxious heat. Conclusion: These data suggest that reserpine in high concentration tested and consequently the dopamine can influence the ANH behavior in D. melanogaster.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: nociception; behavior; dopamine.

ESSENTIAL OILS OF Lippia alba AND Aloysia triphylla ARE ANSIOLITICAL WITHOUT INDUCING AVERSIVENESS IN FISH

ESSENTIAL OILS OF Lippia alba AND Aloysia triphylla ARE ANSIOLITICAL WITHOUT INDUCING AVERSIVENESS IN FISH

Guerino Bandeira Junior 1 , Murilo Sander de Abreu 2 , João Gabriel Santos da Rosa 2 , Leonardo José Gil Barcellos 2 , Bernardo Baldisserotto 2

1 Veterinary Medicine course, Universidade Federal de Santa Maria. 2 Graduate Program in Pharmacology, Universidade Federal de Santa Maria.

Introduction: Essential oils (EOs) from Lippia alba and Aloysia triphylla are known for their anesthetic properties. Most conventional anesthetics used in fish are considered to be aversive to these animals when used at 50% of the anesthetic concentration. Objective: Determine whether these EOs are aversive to fish and whether they possess anxiolytic or anxiogenic properties. Methods: Zebrafish (Danio rerio; n = 60) seven months old (3.6 ± 0.4 cm, 0.58 ± 0.1 g) and five months old silver catfish (Rhamdia quelen; n = 60) juveniles (8 ± 2 cm, 7 ± 2 g) were used. Fish were subjected to a preference test in an apparatus with two water pumps generating two laminar flows without the liquids mixing. The fish could choose to remain on the side containing the substance (EO from L. alba or A. triphylla) or the side containing pure water. In this apparatus, fish were individually filmed for 2 min and 30 s in the pre-flow period (pumps off) and for 2 min and 30 s in the flow period (pumps on). There was a control with water on both sides of the apparatus and a positive (aversive) control with pH 3, obtained with the addition of trichloroacetic acid in the water, on one side of the apparatus. The fish pre-exposed to the substances were immediately transported to the aquarium of experiment 2 (new tank test), where they remained in pure water and were filmed for 6 min. The videos were analyzed using ANY-maze® software. Results: During the flow period, silver catfish and zebrafish remained longer on the side containing water relative to the side containing pH 3 solution (positive control), demonstrating an aversion behavior. The other groups showed no significant difference in length of stay on either side. Silver catfish and zebrafish pre-exposed to the two EOs remained longer in the upper zone of the aquarium, demonstrating an anxiolytic effect. Silver catfish pre-exposed to the L. alba EO demonstrated higher values of distance traveled, mean speed, crossings and entries in the upper zone compared to the control group. Zebrafish pre-exposed to the two EOs showed higher values of distance traveled, mean speed, crossings and entries in the upper zone compared to the control group. Conclusion: EOs of L. alba and A. triphylla, when used at 50% of the anesthetic concentration, have anxiolytic effect in zebrafish and silver catfish, without altering the locomotor patterns nor inducing aversive behavior.

Financial Support: CNPq.

Key words: anesthetics; phytotherapics; anxiolytic.

EXPLORATORY ACTIVITY IS NOT INCREASED AFTER RECURRENT CONCUSSION IN YOUNG RATS Gustavo Cassol 1 ,

EXPLORATORY ACTIVITY IS NOT INCREASED AFTER RECURRENT CONCUSSION IN YOUNG RATS

Gustavo Cassol 1 , Caroline Brandão Quines², Luiz Fernando Freire Royes 12

1 Programa de Pós-Graduação em Educação Física, Universidade Federal de Santa Maria.

2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: By definition, traumatic brain injury (TBI) is classified as a change in brain function caused by the application of an external force and represents a global public health problem. In this context, recurrent and low to medium gravity TBIs are called concussions. As a consequence of these concussions, some behavioral changes may occur due to the unrestricted movement of the head that causes the brain to move within the skull. Objective: The aim of the present study was to evaluate the exploratory activity of young rats submitted to recurrent concussions. Methods: Wistar rats were used at thirty-five days. The animals were submitted to seven concussions with intervals of twenty four hours each. Weight Drop was used as the TCE model. Seventy-two hours after the last concussion, the animals were submitted to the open field test, the parameters of crosses and elevations made by the animal were evaluated. Thirty-two days after the last concussion, the animals were reevaluated with the same test. The animals were kept in a light / dark cycle of 12 hours with water and food ad libitum. The protocol was approved by the Ethics Committee of the Federal University of Santa Maria through the process number 3740020616. Results: The results obtained show that the concussions did not cause statistical differences in the cross-over parameter, nor in elevations between the control and concussion groups. Conclusion: These data suggest than our recurrent concussion model did not induce an increase in exploratory activity in young rats.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: concussion; behavior; mild traumatic brain injury.

SEIZURE-LIKE BEHAVIOR INDUCED BY PTZ IN THE COCKROACH Nauphoeta cinerea Jéssica C. Alvim 1 ,

SEIZURE-LIKE BEHAVIOR INDUCED BY PTZ IN THE COCKROACH Nauphoeta cinerea

Jéssica C. Alvim 1 , Cláudia S. Oliveira 2 , João Batista T. Rocha 1,2

1 Departamento de Bioquímica e Biologia Molecular, CCNE, Universidade Federal de Santa Maria. 2 Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica, Universidade Federal de Santa Maria.

Introduction: Pentylenetetrazol (PTZ) is a classic convulsant widely used in the search for novel anticonvulsants. Studies demonstrated that PTZ induces seizures in experimental models like zebrafish and rats, but, to our knowledge there is no studies about the interaction of PTZ with cockroaches. Therefore, it is important to evaluate the cockroaches behavior after exposure to PTZ. Objective: The aim of this study was to evaluate cockroaches behavior after PTZ exposure to search for alternative models to study seizure-like behavior. Methods: The cockroaches were separated into six groups: Saline 1 (1 µl/cockroach), Saline 2 (2 µl/cockroach), DMSO 1 (1 µl/cockroach), DMSO 2 (2 µl/cockroach), PTZ 1 (1 µmol/lµL/cockroach) and PTZ 2 (2 µmol/2µl/cockroach). The drug was injected into the cockroaches head after cryoanaesthesia (10 minutes in the ice). Thirty cockroaches were used in each group and they were observed during 60 minutes. Parameters such as normal behavior, erratic movements, seizure-like behavior and loss of posture were evaluated. Mortality of cockroaches was followed for ten days. Results: No significant difference was observed in mortality of cockroaches. None of the cockroaches exposed to Saline (1 and 2) and DMSO (1 and 2) exhibited seizures-like behavior. However, the cockroaches from DMSO 2 exhibited erratic movements. Regarding to PTZ exposure, 80% and 100% of the cocroaches exposed to PTZ 1 and PTZ 2, respectively, exhibited seizures, the main sign was loss of posture and erratic movements. Conclusion: These results suggest that the two PTZ concentrations tested caused a seizure-like behavior and further studies are needed to determine the participation of GABA receptors in the process by administration of GABA agonists.

Financial Support: CNPq, CAPES, FAPERGS.

Key words: cockroaches; behavior; seizures.

ACUTE AND CHRONIC EXPOSURE TO ALARM SUBSTANCE: INVOLVEMENT ON AGGRESSIVE CONTEXT OF TWO ZEBRAFISH STRAINS

ACUTE AND CHRONIC EXPOSURE TO ALARM SUBSTANCE:

INVOLVEMENT ON AGGRESSIVE CONTEXT OF TWO ZEBRAFISH STRAINS

Julia Canzian 1 , Vanessa Andreatta Quadros 1,2 , Talise Ellwanger Müller 1,2 , Fabiano Vargas da Costa 1 , Denis Broock Rosemberg 1,2

1 Laboratory of Experimental Neuropsychobiology, Department of Biochemistrand Molecular Biology, Natural and Exact Sciences Center, Federal University of Santa Maria. 1000 Roraima Avenue, Santa Maria, RS, 97105-900, Brazil. 2 Graduate Program in Biological Sciences: Toxicological Biochemistry, Federal University of Santa Maria. 1000 Roraima Avenue, Santa Maria, RS, 97105-900, Brazil.

Introduction: Aggression is associated with an intrinsic modulation of different neurochemical parameters, influencing the release of neurotransmitters and hormones, enzyme activity, and gene expression. Recent data with different zebrafish strains show the importance of performing neurobehavioral studies to comprehend the mechanisms involved in human diseases as well as on aggression. Both WT and leo strains present behavioral differences, in which leo has pronounced defensive responses, suggesting increased anxiety-like behavior. These behavioral responses can be modulated with the "alarm substance" (AS) or “schreckstoff” released into the water when the animal skin is damaged. Objective: We evaluated the agonistic profile of two strains of zebrafish (WT and leo) after acute and chronic exposure to AS. Materials e Methods: The AS was extracted by damaging epithelial cells using donor fish previously euthanized into 10 mL distilled water. Fish were individually exposed to AS for 5 min (acute) or during 7 days, 5 min once daily (chronic protocol). After, fish were placed in the test tank containing an inclined mirror on the outside part of the apparatus and the agonistic behaviors were recorded for a single session of 6 min. The tank was virtually divided into three areas related to their proximity to mirror and the following endpoints were analyzed: distance traveled and absolute turn angle (locomotors parameters), number and time of aggressive episodes, and latency to attack the mirror (agonistic behaviors). Results: The acute exposure to AS significantly increased the number and time of aggressive episodes in leo in comparison to its respective control. The chronic exposure to AS decreased the distance traveled and the time of aggressive display in WT in relation to its control group. AS did not alter the other behavioral endpoints measured. Conclusion: We report that the effects of AS on aggressive behavior depend on strain and time of exposure.