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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

Anfiteatro do Colgio Politcnico


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 Committee
Prof. Allan V. Kalueff (St Peterburg State University, Russia, ZNRC Chair)
ISBS President
Ana Lcia 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
Cludia 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 Vincius 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. Mller, 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 Sciences: Toxicological Biochemistry
(UFSM)
Graduation Program in Pharmacology (UFSM)
Study English
LabSul
Sorrisos 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 nonmammalian 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
Paulo A. Burmann (Dean, UFSM)
Flix 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
ROUND TABLE CONFERENCE II: TRANSLATIONAL MODELS OF
NEUROPSYCHIATRIC DISORDERS
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 paclitaxelinduced 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: Cludia S. de Oliveira (UFSM) - Mercury: interaction with Selenium
Speaker 3: Joo 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
19:00 FAREWELL CONFRATERNIZATION (SORRISO'S GASTRONOMIA)

FINAL PROGRAM

ABSTRACTS

SESSION: TOXICOLOGICAL BIOCHEMISTRY

PROTECTIVE EFFECTS OF QUERCETIN ON REACTIVE OXYGEN


SPECIES IN RATS SUBMITTED TO EXPERIMENTAL ADJUVANT
ARTHRITIS MODEL
Alessandra G. Manzoni1, Renata S. Pereira1,2, Lvia G. Castilhos1,2, Juliana Sorraila3,
Lara Vargas Becker1,3, Karine Lanes Silveira1,2, Daniela B. R. Leal1,2,3
1

Laboratrio de Imunobiologia Experimental e Aplicada, Universidade Federal de Santa Maria.


Programa de Ps-Graduao em Cincias Farmacuticas:Anlises Clnicas, Universidade Federal de Santa
Maria.
3
Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
2

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 antiinflammatory 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 Kegler1, Eduardo T. Pascotini1, Josi Arend2, Michele R. Fighera1,2
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de


Santa Maria.
2
Programa de Ps-Graduao 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- (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- 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 COCKROACHES
Allan P. Leal1, Raquel O. Soares1, Ana Paula A. Perin1, Bruna T. Borges1, Barbara A.
B. Ogata1, Etiely Karnopp1, Maria E. Rosa1, Yuri C. Barreto1, Ana P. Zanatta1, Tiago
G. dos Santos3, Chiara Valsecchi1, Lcia Vinad1, Christon Andr Dal Belo1,2
1

Laboratrio de Neurobiologia e Toxinologia, (LANETOX), Universidade Federal do Pampa. 2Programa de


Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de Santa Maria.
3
Laboratrio 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 100g/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 8g/200l)
on CSIHP was dose-dependent with a maximum decrease of heart rates (504 beats/min) for the highest
concentration, in 30min (n=9, p<0.05). MERIV (2, 10 and 20 g/g animal weight) significantly increased
grooming activity that was prevented by the previous treatment with phentolamine (0.1g/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. B. Ogata1, Raquel S. Oliveira1, Liane Vargas2, Allan Pinto Leal , Bruna
Trindade Borges, Maria Eduarda Rosa, Yuri Correia Barreto, Ana Paula Zanatta,
Etiely Karnop, Pamela M. Carpes2, Christon Dal Belo1, Lcia Vinad1.
1
2

Laboratrio de Neurobiologia e Toxinologia, Universidade Federal do Pampa, So Gabriel, RS.


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 M, 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 M 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 M 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. Borges1, Allan P. Leal1, Ana P. Zanatta1, Barbara A. B. Ogata1, Etiely
Karnopp1, Maria E. Rosa1, Yuri C. Barreto1, Lcia Vinad 1, Christon Andr Dal
Belo1,2, Patrcia B. Vieira1
1

Laboratrio de Neurobiologia e Toxinologia (LANETOX), Universidade Federal do Pampa.


Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
2

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 l of the test compounds previously
dissolved in physiological solution. The crude extract of M. rufula (CEMR) was administered at 50, 100 and
200 g per animal and the methanolic (MF) and aqueous (AF) fractions were administrated at 10, 20 and 40
g 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 g/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 Piccoli1, Cludia Sirlene de Oliveira1, Fernanda Dvila da Silva1,
Jssica da Costa Alvim2, Joo Batista Teixeira da Rocha1,2
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de


Santa Maria.
2
Departamento de Bioqumica 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 (Hg0), inorganic (Hg2+), 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 g/g of food) during three months; 2) Cockroaches were fed with different concentrations of MeHg
(0, 2.5, 5, 25 and 100 g/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 OF Drosophila melanogaster
Cynthia Camila Ziech1, Eduardo da Rosa vila1, Nathane Rosa Rodrigues2, Giulianna
Echeverria Macedo1, Gustavo Felipe da Silva1, Karen Kich Gomes1, Jessica Ferreira
Rodrigues, Jeferson Luis Franco1, Thas Posser1
1

Oxidative Stress and Cell Signaling Research Group (GPEOSCEL), Universidade Federal do Pampa,
Campus So Gabriel, 97300-000 So Gabriel, RS, Brazil;
2
Departamento de Qumica, Programa de Ps Graduao em Bioqumica Toxicolgica, 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 Parkinsons 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
Dbora Gonalves1, Nelson R. de Carvalho2, Aline Courtes1, Pamela Carvalho1, Ingrid
Kich1, Flix A. Soares1, Thais Poser2, Jeferson L. Franco2.
1

Departamento de Bioqumica e Biologia Molecular, Centro de Cincias Naturais e Exatas, Universidade


Federal de Santa Maria, Campus UFSM, Santa Maria, RS, Brazil. 2Campus So Gabriel, Universidade
Federal do Pampa, So 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 oC using DatLab 4.0
software (Oroboros Inc., Austria), with continuous stirring at 750 rpm. HRR analyses coupled to substrateuncoupler-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
Keulss 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 Karnopp1, Douglas S. Santos1,3, Raquel S. Oliveira1, Allan P. Leal1, Ana P.
Zanatta1, Barbara A. B. Ogata1, Bruna T. Borges1, Maria E. Rosa1, Lucas F. Maciel1,
Yuri C. Barreto1, Lcia Vinad1, Jaderson C. Costa2, Paulo M. Pinto4 Christon A. Dal
Belo1,2,3
1

Laboratrio de Neurobiologia e Toxinologia, (LANETOX), Universidade Federal do Pampa. 2Instituto do


Crebro (INSCER), Pontifcia Universidade Catlica do Rio Grande do Sul. 3Programa de Pos-Graduao em
Cincias Biolgicas: Bioqumica Toxicolgica. 4Laboratrio de Protemica 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 Ca2+ 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 g/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 Ca2+
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 Obetine Baptista, Leticia Priscilla Arantes, Alisson Rodrigues Vinholes,
Aline Frazen da Silva, Danela de Freitas Camara, Marina Lopes Machado, Tssia
Limana da Silveira, Thayanara Cruz da Silva, Flix Alexandre Antunes Soares.
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, 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 (5hydroxy-1,4-naphthoquinone) 200 M, 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 M and 3 at 2M. 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
Gustavo H. Weimer2, Aline M. B. do Amaral1,2, Aline da S. Faria2, Carolina L.
Gressler2, Dionatan de Pellegrin2, Gracielle da S. Flix2, Letcia K. de Moura2, Kamila
E. Menezes2, Rafaela R. M. de Souza2, Vania Lucia Loro1,2
1
2

Programa de Ps-Graduao em Biodiversidade Animal, Universidade Federal de Santa Maria


Laboratrio de Toxicologia Aqutica, 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 melanogaster
Jssica Ferreira Rodrigues1, Karen Kich Gomes1, Nathane Rosa Rodrigues2, Giulianna
Echeverria Macedo1, Illana Kemerich Martins1, Cynthia Camila Ziech1, 1Thas Posser,
Patrcia de Brum Vieira1, Jeferson Luis Franco1.
1

Oxidative Stress and Cell Signaling Research Group (GPOSCEL), Universidade Federal do Pampa, Campus
So Gabriel, 97300-000 So Gabriel, RS, Brazil;
2
Departamento de Qumica, Programa de Ps Graduao em Bioqumica Toxicolgica, 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 FolinCiocalteau 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 M
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
melanogaster
Karen Kich Gomes1, Giulianna Echeverria Macedo1, Nathane Rosa Rodrigues1, Jssica
Ferreira Rodrigues1, Cynthia Camila Ziech1, Jeferson Luis Franco1, Thas Posser1.
1

Grupo de Pesquisa Estresse Oxidativo e Sinalizao Celular: Universidade Federal do pampa; Campus So
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
Letcia Kuhn de Moura1, Jossiele Wesz Leitemperger2, Ana Carolina Gressler1, Talise
Ellwanger Mller2, Bibiana Silveira Moraes3 Vania Lucia Loro2,3
1

Graduanda em Zootecnia, Universidade Federal de Santa Maria.


Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
3
Programa de Ps-Graduao em Cincias Biolgicas: Biodiversidade Animal, Universidade Federal de
Santa Maria.
2

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 (H2O2) 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 TrisHCl 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
Lvia F. Davila1, Vernica T. Dias1, Luciana T. Vey2, Karine Roversi1,Laura H.
Milanese1, Fabola Trevizol1, Marilise E. Brger1,2
1

Programa de Ps-Graduao em Farmacologia, Universidade Federal de Santa Maria.


Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
2

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
Lvia G. Castilhos1,2, Fatima H. Abdalla2, Jamile F. Gonalves2, Fernanda L. Cabral1,
Alessandra G. Manzoni2, Daniela B.R. Leal1,2
1

Programa de Ps-Graduao em Cincias Farmacuticas, Universidade Federal de Santa Maria.


Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
2

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 cats 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-MazeTM 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; cats claw; acetylcholinesterase; senescence.

TOXICITY OF AGROCHEMICAL MANCOZEB IN EMBRYO


DEVELOPMENT AND BEHAVIOR OF ZEBRAFISH
Luana Paganotto Leandro1, Dennis Guilherme da Costa Silva2, Andressa Rubim
Lopes1, Illana Kemmerich Martins2, Lucia Emanueli Schimith3, Nelson Rodrigues de
Carvalho2, Mauro Eugnio Medina Nunes4, Thais Posser5, Jeferson Luis Franco5.
1

Cincias Biolgicas Bacharelado, Universidade Federal do Pampa; So Gabriel, Rio Grande do Sul;
Programa de Ps-Graduao em Cincias Biolgicas da Universidade Federal do Pampa; So Gabriel, Rio
Grande do Sul;
3
Biotecnologia, Universidade Federal do Pampa; So Gabriel, Rio Grande do Sul;
4
Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica da Universidade Federal de
Santa Maria; Santa Maria, Rio Grande do Sul;
5
Universidade Federal do Pampa; So Gabriel, Rio Grande do Sul.
2

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, 500.S cm-1, 28C). Embryos, 3-4 hours post fertilization (hpf) were pre-exposed during 1h to the
antioxidant N-acetylcysteine (NAC; 750M). 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-MazeTM 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 sensorymotor 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. Pretreatment with NAC blocked all morphological changes, locomotor /sensory responses and decreased MZinduced 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. Schimith1, Mauro E.M. Nunes2, Dennis G.C. Silva3, Nelson Rodrigues de
Carvalho3, Andressa R. Lopes4, Luana P. Leandro4, Thais Posser3, Jeferson L. Franco5
1

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


Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica da Universidade Federal
de Santa Maria, Santa Maria, Rio Grande do Sul.
3
Programa de Ps-Graduao em Cincias Biolgicas da Universidade Federal do Pampa. So Gabriel, Rio
Grande do Sul.
4
Cincias Biolgicas Bacharelado, Universidade Federal do Pampa. So Gabriel, Rio Grande do Sul.
5
Orientador. Universidade Federal do Pampa.
2

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 g.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-MazeTM
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 g.L-1). After, two
concentrations were chosen, one lower and one higher than LC50 (25 and 50 g.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. Braga1, Ben Hur M. Mussulini2, Emerson S. Silva2, Gabriela Lazzarotto2,
Diogo L. Oliveira2, Diogo O. Souza2, Joo Batista T. Rocha1
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de


Santa Maria.
2
Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica, 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 O2 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 3rd 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. Rosa1, Douglas S. dos Santos3, Ana P. Zanatta1, Bruna T. Borges1, Barbara
A. B. Ogata1, Lucas Maciel1, Yuri C. Barreto1, Allan P. Leal1, Etiely Karnopp1, Lcia
Vinad1, Ernani Pinto2, Christon A. Dal Belo1,3
1

Laboratrio de Neurobiologia e Toxinologia (LANETOX), Universidade Federal do Pampa.


Departamento de Anlises Clnicas Toxicolgicas da Faculdade de Farmcia, Universidade de So Paulo.
3
Programa de Ps-graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
2

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 So 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 n 043/2015). Results:
ECATAS (2.5, 5 and 50g/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 8g/ml) induced a maximum decreased in the cell viability (4g/ml), compared to
the HEPES control (p <0.05, n=6). Using BCMP and CNP, ECATAS (2, 4 and 8g/ml) and (5, 25 and 50 g/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 S. C. dos Santos 1,2, Tase F. Pedroso 2, Maria Ester Pereira 2,3
1

Centro de Cincias da Sade (CCS)-Federal University of Santa Maria, Santa Maria, RS, Brazil.
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.
2

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 Ac2PbWistar rats. Methods: The animals were treated as follows: received saline, zinc chloride
(ZnCl2 27 mg/kg), N-acetylcysteine (NAC 5 mg/kg) or ZnCl2 + NAC day 3 to 7; lead acetate (Ac2Pb 1th to
12th 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 Ac2Pb 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
Marina Lopes Machado, Letcia Priscilla Arantes, Thayanara Cruz da Silva, Tssia
Limana da Silveira, Daniela de Freitas Cmara, Fabiane Bicca Obetine Baptista,
Flix Alexandre Antunes Soares
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, 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 Viana1, Andressa Palharini Machado1, Mara Lisiane Tissot Squalli
Houssaini2, Ilaine Teresinha Seibel Gehrke2
1

Graduao em Cincias Biolgicas- Bacharelado, Universidade Regional do Noroeste do Estado do Rio


Grande do Sul.
2
Quadro docente do departamento de Cincias 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
Naili Schiefelbein Souto1, Micheli Dassi1, Ana Claudia Monteiro Braga2, rica Furlan
da Rosa2, Ana Flvia Furian1,2
1
2

Programa de Ps-Graduao em Cincia e Tecnologia em Alimentos, Universidade Federal de Santa Maria.


Programa de Ps-Graduao 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
proteins, RNA and DNA. 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 g/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 3nitrotyrosine, 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 (PKCSer957). 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 Ser957/PKC ). Conclusion: AFB1 was able to cause neurochemical
alterations. These results reinforce that monitoring mycotoxins levels in food are essential to guarantee food
security.
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. Nogara1, Alcindo Busanello2, Nilda B. V. Barbosa1, Roselei Fachinetto1,2,
Joo B. T. Rocha1
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de


Santa Maria. 2Programa de Ps-Graduao 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 - 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 - 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 PROOXIDANTS.


Renata G. Budel1, Larissa F. Schaffer2, Roselei Fachinetto3, Luis R. Peroza2
1

Programa de Ps-Graduao em Nanocincias, Centro Universitrio Franciscano.


Docentes do curso de Biomedicina, Centro Universitrio Franciscano.
3
Programa de Ps-Graduao em Farmacologia, Universidade Federal de Santa Maria.
2

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 37C with pro-oxidants and
in the presence or absence of different concentrations of isoflavones (equivalent to 25, 50, 100, 200 and 300
g/mL of isoflavones). As pro-oxidant agents, sodium nitroprusside (NPS 5M) and Fe2+/EDTA (100 M/100
M) 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 Fe2+/EDTA complex increased TBARS in brain
homogenates of the rat. Isoflavones reduced this increase in a concentration-dependent manner with an IC50 of
71.39 2.01g/ml to NPS and 124.5 8.13 g/ml to Fe2+/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 dos Santos 1,3, Jamile S. Bernardi 2, Maria Ester Pereira 2,3.
1

Centro de Cincias da Sade (CCS) - UFSM Universidade Federal de Santa Maria, Santa Maria, RS, Brasil.
Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria, Santa Maria, RS, Brasil.
3
Departamento de Bioqumica e Biologia Molecular, Centro de Cincias Naturais e Exatas (CCNE).
2

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 10th 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 Marion 1,2, Jamile S. Bernardi 2, Maria Ester Pereira 2,3.
1

Centro de Cincias da Sade (CCS) - UFSM Universidade Federal de Santa Maria, Santa Maria, RS, Brasil.
Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria, Santa Maria, RS, Brasil.
3
Departamento de Bioqumica e Biologia Molecular, Centro de Cincias Naturais e Exatas (CCNE).
2

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 27th 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 27th to the 30th 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 Mller1, Mauro Medina Nunes1, Charlene Cavalheiro Menezes1,
Flavia Vestena Stefanello1, Vania Lucia Loro1, Denis Broock Rosemberg1
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, 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 Parkinsons 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 Na2SeO3 dietary on biochemical parameters of zebrafish exposed to PQ. Methods: Initially, adult zebrafish
of both sexes were divided into two groups: control diet (without Na2SeO3) and SE diet (1.0 mg/Kg Na2SeO3)
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: Na2SeO3 dietary prevented the increase of carbonylated
protein (CP), reactive oxygen species (ROS) and nitrite/nitrate (NOx) levels, as well as the decrease in nonprotein thiols (NPSH) levels caused by PQ. Regarding the antioxidant enzymatic defenses, Na2SeO3 dietary
prevented the increase in catalase (CAT) and glutathione peroxidase (GPx) activities induced by PQ.
Conclusion: Altogether, Na2SeO3 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; Parkinsons disease.

QUINOLIC ACID INDUCES TOXICITY IN Caenorhabditis elegans


Tssia L. da Silveira, Daniele C. Zamberlan, Marina L. Machado, Thayanara C.
Silva, Leticia P. Arantes Fabiane B. O. Baptista, Felix A. A. Soares
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, 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 oC for 1
hour. The analyses included evaluation of QAs 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 DAF16/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 C. Barreto1,2, Alan P. Leal1, Raquel S. Oliveira1, Ana P. Zanatta1, Barbara A. B.
Ogata1, Bruna T. Borges1, Etiely Karnopp1,4, Maria E. Rosa1, Lucas F. Maciel1,
Douglas S. Santos3, Tiago G. dos Santos2, Christon A. Dal Belo1,3, Lucia Vinad1.
1

Laboratrio de Neurobiologia e Toxinologia (LANETOX), Universidade Federal do Pampa, So Gabriel,


Brasil.
2
Laboratrio de Estudos do Bioma Pampa (LEBIP), Universidade Federal do Pampa, So Gabriel, Brasil.
3
Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, 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 n 037/2012). Results: When MERIV (5 and 10 g/mL) was incubated on BCMP there was
an increase in the muscle twitches followed by an irreversible neuromuscular blockade, in 120min
recordings (6015% and 466%, p<0.05, n=5, respectively). MERIV (5, 10 and 20 g/mL)
significantly inhibited AChE activity (235%, 565% and 526%, 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 (276% and 19 4%, p<0.05, n=5, respectively). MERIV
(5, 10 and 20 g/mL) increased the mouse brain cell viability at the lowest concentration (3610%,
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.
Financial Support: Edital de Toxinologia CAPES 063/2010, PRONEM /FAPERGS/CNPq
003/2011.
Key words: Toad venom; Acetylcholinesterase; neurotoxicity.

FINAL PROGRAM

ABSTRACTS

SESSION: NEUROPSYCHOPHARMACOLOGY

ANESTHETIC ACTIVITY OF THYMOL AND CARVACROL IN SILVER


CATFISH (Rhamdia quelen).
Adriane E. Bianchini1, Jessyka A. da Cunha1, Isabel C. M. Brusque2, Guerino Bandeira
Junior2, Bernardo Baldisserotto1
1
2

Pharmacology Graduation Program, Universidade Federal de Santa Maria.


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. Almeida1, Berta M. Heinzmann2, Adalberto L. Val3, Bernardo Baldisserotto1
1

Animal Biodiversity Graduation Program, Universidade Federal de Santa Maria.


Departmento of Industrial Pharmacy, Universidade Federal de Santa Maria.
3
Laboratrio de Ecofisiologia e Evoluo Molecular, Instituto Nacional de Pesquisas da Amaznia
2

Introduction: Practices performed in aquaculture as eggs extrusion, biometry and transport expose
fish to intense stress, resulting in damage to the animals 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 L L-1 and 50, 100 or 200 L 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 L 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 L 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 L 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 Ceretta1, Catiuscia Molz de Freitas2, Larissa Finger Schaffer1,
Mariana Maikli Dotto3, Jeane Binotto Reinheimer3, Elizete de Moraes Reis1, Roselei
Fachinetto1,2
1

Programa de Ps-Graduao em Farmacologia, Universidade Federal de Santa Maria.


Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
3
Curso de Farmcia, Universidade Federal de Santa Maria, Santa Maria.
2

Introduction: Tardive dyskinesia is a common side effect in patients with long-term typical antipsychotic
medication and vacuous chewing movements 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 28th 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 meanstandard error
of the mean (SEM). Statistical analysis of data was carried out by one-way analysis of variance (ANOVA)
followed by Tukeys 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. Fontana1, Daniele L. Meinerz2, Nathana J. Mezzomo3, Luiz Vincius C.
Rosa1, Vanessa A. Quadros1, Denis B. Rosemberg1
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de


Santa Maria.
2
Laboratrio de Neuropsicobiologia Experimental, Universidade Federal de Santa Maria.
3
Programa de Ps-Graduao 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 GABAA 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 MODEL IN MICE
Evelyne S. Brum1, Gabriela Beck2, Sara M. Oliveira3
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de


Santa Maria.
2
Laboratrio de Neurotoxicidade e Psicofarmacologia, Universidade Federal de Santa Maria.
3
Departamento de Bioqumica 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 CEUAUFSM (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 L 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 L) 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 83.45.1% at 4 h after treatment. In addition, mice treated with Eta were capable of
reduce (61.912.4%) 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. da Cruz1; Fernanda Windmoller Brendler2; Andria Kusiack Gollo2;
Mauricio Borges da Rosa3; Emanuelle Bortolotto Degregori2
1

Docente do Curso de Medicina Veterinria, Departamento de Estudos Agrrios, Universidade Regional do


Noroeste do Estado do Rio Grande do Sul.
2
Aluno de Graduao em Medicina Veterinria, Universidade Regional do Noroeste do Estado do Rio Grande
do Sul.
3
Mdico Veterinrio, Hospital Veterinrio, Departamento de Estudos Agrrios, 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 () receptors
exerting analgesic effect by reducing pain neurotransmitters. Methadone is a 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 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
alpha2 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 (16,13,9Kg) 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 oC 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 oC 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-UNIJU
Key words: epidural; NMDA receptor; opioids.

EFFECTS OF AMPH-ISOTERAPIC ON AMPH DEPENDENCE


PARAMETERS
Hecson J. Segat1, Vincia G. Metz3, Higor Z. Rosa3, Raquel C. S. Barcelos2, Marilise
E. Burger1,2,3
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de


Santa Maria.
2
Programa de Ps-Graduao em Farmacologia, Universidade Federal de Santa Maria.
3
Centro de Cincias da Sade, 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 10GL) 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 RATS
Laura H. Milanesi1, Karine Roversi1, Caren D. Antoniazzi1, Hecson J. Segat2, Maikel
Kronbauer1, Lvia F. Dvila1, Veronica T. Dias1, Marcel H.M. Sari2, Raquel C.S.
Barcelos1, Fabola Trevizol1, Marilise E. Burger12*
1

Programa de Ps-Graduao em Farmacologia, Universidade Federal de Santa Maria.


Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
2

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 (n041235/2016). 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. Bhler1, C.G. Zinn1, N. Clairis2, L.E. Cavalcante1, C.R.G. Furini1, I. Izquierdo1, J.C.
Myskiw1
1

Memory Center, Brain Institute of Rio Grande do Sul, Pontifical Catholic University of Rio Grande do Sul
(PUCRS) Porto Alegre RS, Brazil.
2
Dpartement 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 L) or into BLA (0.5 L) 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, -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, -adrenoceptor antagonist, timolol (1.0 g/side), p<0.05 (Veh-N vs. TimN), 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 RATS
Luciana T. Vey1, Higor Z. Rosa2, Vincia G. Metz2, Vernica T. Dias3, Lvia F.
Davila3, Marilise E. Burger1,2,3
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de


Santa Maria.
2
Departamento de Fisiologia e Farmacologia, UFSM-RS, Brasil
3
Programa de Ps-Graduao 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 rats 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 gestS 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 FOLLOW-UP STUDY
Lucas Maciel1, Fernanda Barbisan2, Ivana B. Manica da Cruz2, Tlis de Oliveira Silva2
1

Laboratrio de Neurobiologia e Toxinologia, (LANETOX), Universidade Federal do Pampa. 2Programa de


Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, 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 Maus city, AM-Brazil during a period
of 5-years. Elderly (> 60 years old) of the Family Health Strategy Program (FHS-SUS) of Maus
(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 GUARAN CHEMICAL MATRIX COMPOUNDS ON THE


REVERSAL OF ANTIPSYCHOTIC DRUG-INDUCED INFLAMMATION IN
RAW 264.7 LINEAGE MACROPHAGES
Moiss Henrique Mastella1, Fernanda Barbisan2, Vernica Farina Azzolin2, Pedro
Antonio Schmidt Prado-Lima3, Gustavo Cardenas4, Euler Ribeiro5, Cibele Ferreira
Teixeira2, Vitor Bueno4, Ivana Beatrice Mnica da Cruz2, Emilio Ivo da Cruz Jung2.
1

Curso de Cincias Biolgicas, Universidade Federal de Santa Maria;


Programa de Ps-Graduao em Farmacologia, Universidade Federal de Santa Maria.
3
Instituto do Crebro, Pontifcia Universidade Catlica 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).
2

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
guaran (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 guaran. 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 guaran 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- [4,5dimethylthiazol-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 guaran
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 guaran 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: Guaran; inflammation; antipsychotic.

EFFECT OF SUBCHRONIC TREATMENT OF CYCLOOXYGENASE-2


INHIBITORS ON PENTYLENETETRAZOL (PTZ)-INDUCED SEIZURES IN
MICE
Paula Michelotti1, Fernanda Rossatto Temp1, Joseane Righes Marafiga1, Ana Cludia
Jesse1, Laura Hautrive Milanesi1 and Carlos Fernando Mello1
1

Programa de Ps-Graduao 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 AMPHETAMINECONDITIONED PLACE PREFERENCE IN YOUNG ADULT RATS
Vernica Tironi Dias1, Higor Zuqueto Rosa3, Lvia Ferraz Davila1, Luciana Taschetto
Vey2, Raquel Cristine Silva Barcelos1, Marilise Escobar Burger1,2,3
1

Programa de Ps-Graduao em Farmacologia, Universidade Federal de Santa Maria.


Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
3
Departamento de Fisiologia e Farmacologia, Universidade Federal de Santa Maria.
2

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
Vincia G. Metz2, Hecson J. Segat1, Marilise Escobar Brger1
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, 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

MODULATION OF GROOMING BEHAVIOR INDUCED BY THE UREASE


OF Canavalia ensiformis (JBU) IN Nauphoeta cinerea COCROACHES
Ana Paula Zanatta1, Douglas Silva dos Santos1,2, Carlos Gabriel Moreira de Almeida,
Maria Eduarda Rosa1, Bruna Trindade Borges1, Yuri Correia Barreto1, Lucas Maciel1,
Allan Pinto Leal1, Barbara Akemi Ogata1, Etiely Karnop1, Lcia do Canto Vinad1,
Clia Regina Carlini3, Christon Andr Dal Belo1,2,3
1

Laboratrio de Neurobiologia e Toxininologia (LANETOX), Universidade Federal do Pamp


Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
2

LANEUROTOX, Instituto do Crebro, Pontifcia Universidade Catlica 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 23-26C, 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 (10l) 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 g/g of animal) was
administered, there was a dose-dependent alteration in the grooming activity. Thus, at 1.5 g/g of animal
weight JBU, no alteration in the grooming activity was observed (n = 30). When 3 g/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 g/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 g/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 (Cyperus


esculentus L.) AND WALNUT Tetracarpidium conophorum MLL. ARG.) ON
SEXUAL BEHAVIOR, HORMONAL LEVEL AND ANTIOXIDANT STATUS
IN MALE RATS
Ayodeji Augustine Oabiyi1, 2, 3, Ganiyu Oboh1, Stephen Adeniyi Adefegha1
1

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


Afe Babalola University, Ado Ekiti. Ekiti State. Nigeria.
3
Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
2

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 Mll. 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
Mll. 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 IN Caenorhabditis elegans
Daniela Freitas Cmara, Marina Lopes Machado, Letcia Priscilla Arantes,
Thayanara Cruz da Silva, Tssia Limana da Silveira, Fabiane Bicca Obetine Baptista,
Alisson Vinholes, Flix Alexandre Antunes Soares.
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, 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-((((4methoxyphenyl) 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 M 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 M 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), unc46 (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-47dependent 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 Silva1, Marcos Martins Braga2, Maria Elisa Calcagnotto2
1

Biochemistry Department, NNNESP Lab, UFRGS, Porto Alegre, RS, Brazil;


Programa de Ps-Graduao em Bioqumica Toxicolgica, Departamento de Bioqumica e Biologia
Molecular, Centro de Cincias Naturais e Exatas, Universidade Federal de Santa Maria, Santa Maria, RS,
Brazil.
2

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 O2 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 O2 levels by addition of nitrogen gas. The
O2 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
rica Furlan da Rosa1, Naili Schiefelbein Souto2, Micheli Dassi2, Ana Claudia
Monteiro Braga1, Ana Flvia Furian1,2
1
2

Programa de Ps-Graduao em Farmacologia, Universidade Federal de Santa Maria


Programa de Ps-Graduao em Cincia 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 g/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 R. Santos1,2, Lucas B. Oliveira2, Lenen Peterson2, Daniel Ardisson-Arajo1,2
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de


Santa Maria.
2
Laboratrio 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 cladeA (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 BY
CHLORPYRIFOS
Fernanda DAvila da Silva1, Marcos Martins Braga1, Joo Batista Teixeira da Rocha1,2
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade


Federal de Santa Maria.
2
Departamento de Bioqumica 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 L/cockroach) and the animals from the
CPF and CPF + P groups were exposed to CPF 3.4 nmol/10 L/cockroach. After 10 minutes, the Control and
CPF groups were treated with saline (10 L/cockroach), and P and CPF + P groups were treated with
pralidoxime 120 nmol/10 L/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.,(IC); Oliveira, Renata S. C.(IC); Sangoi, Larissa G.(IC);
Bordignon, Eduardo M. (IC); Daronco, Luciane S.E. (O)
1
2

Departamento de Educao Fsica Bacharelado, Universidade Federal de Santa Maria.


Departamento de Cincias da Sade, 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 Vestena Stefanello1, Talise Ellwanger Mller1,2, Mauro Medina Nunes2, Denis
Broock Rosemberg1,2, Vania Lucia Loro2
1

Laboratrio de Neuropsicobiologia Experimental, Universidade Federal de Santa Maria.


Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
2

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 Na2SeO3 dietary
prevents behavioral changes triggered by PQ in zebrafish. Methods: In this experiment, the fish were
pretreated with a diet supplemented with 1 mg/kg Na2SeO3 for 21 days. Fish were further subdivided in four
experimental groups: control group (CT), paraquat group (PQ), Na2SeO3 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, Na2SeO3 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, Na2SeO3 prevented the increase in the latency to enter the area closer to conspecifics and the
agonistic behavior of PQ-treated animals, respectively. Conclusion: The Na2SeO3 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; Parkinsons disease.

EFFECT OF RESERPINE ON THE AVOIDANCE OF NOXIOUS HEAT


BEHAVIOR IN Drosophila melanogaster
Getulio N. Bressan1, Roselei Fachinetto1,2
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de


Santa Maria.
2
Programa de Ps-Graduao 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 M) 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 46C. Results: Through TTP was verified that the 46C temperature was harmful to the fly. Flies
treated with 10 and 30 M of reserpine showed impairment of locomotion activity. However, only the
concentration of 30 M 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 M 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
Guerino Bandeira Junior1, Murilo Sander de Abreu2, Joo Gabriel Santos da Rosa2,
Leonardo Jos Gil Barcellos2, Bernardo Baldisserotto2
1
2

Veterinary Medicine course, Universidade Federal de Santa Maria.


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 Cassol1, Caroline Brando Quines, Luiz Fernando Freire Royes12
1

Programa de Ps-Graduao em Educao Fsica, Universidade Federal de Santa Maria.


Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
2

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
Jssica C. Alvim1, Cludia S. Oliveira2, Joo Batista T. Rocha1,2
1

Departamento de Bioqumica e Biologia Molecular, CCNE, Universidade Federal de Santa Maria.


Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
2

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 l/cockroach), Saline 2 (2
l/cockroach), DMSO 1 (1 l/cockroach), DMSO 2 (2 l/cockroach), PTZ 1 (1 mol/lL/cockroach) and PTZ
2 (2 mol/2l/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
Julia Canzian1, Vanessa Andreatta Quadros1,2, Talise Ellwanger Mller1,2, Fabiano
Vargas da Costa1, Denis Broock Rosemberg1,2
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.
Financial Support: FAPERGS, CNPq, CAPES.
Key words: aggression; agonistic behavior; schreckstoff.

PERSONALITY ASPECTS AND NEUROBIOLOGY OF PEOPLE WITH


PSYCHOPATHY
Sangoi, Larissa G.; Bordignon, Eduardo M.; Oliveira, Renata S. C.; Fernandes,
Carlos A. O. R.; Costa, Gabriela M.
Departamento de Neuropsiquiatria, Universidade Federal de Santa Maria.
Departamento de Educao Fsica Bacharelado, Universidade Federal de Santa Maria.
Introduction: The psychopathy corresponds to a personality disorder manifested by an increased risk of
antisocial behavior and emotional deficits, involving a raising of aggressiveness, a reduction of empathy and
guilty, an absence of remorse and a smaller harm avoidance. Unfortunately, despite the serious impairments
to social interaction, just a little is known about this disturbs underlying neurobiology, due to the scarcity of
the existent studies and the difficulty to form groups of patients for research. Objectives: To analyze the profile
of behaviors between people with psychopathy and to know the recent scientific production about the diseases
neurobiological bases. Methodology: A systematic review of the last five years literature was developed,
from PubMed, in which the key words Antisocial Personality Disorder, Neurobiology and its synonyms
were crossed. In total, 25 articles were found, being chosen 13 of these, without idiom restriction. Articles with
different thematic approach, including those that focused in the influence of sociocultural factors, were
excluded. Results: It is consensual among authors that the impairments to social interaction come from an
apparent disconnection between fronto-limbic structures, pointing out the hypofunction of amygdala in
reinforcement learning and of ventromedial frontal and cingulated anterior cortex in value representation.
These alterations justify common traces found in individuals with psychopathy, like absence of anxiety,
depression and guilty, inability of express affection and difficulty in recognize what can be important for
others. Interestingly, electroencephalogram (EEG) exams showed differences in the electrical activity of
patients brain, compared to controls. Sums up to that a reduction in the serotonin function (especially in the
impulsiveness-aggressiveness) and a dysfunction in the endogenous opioids system. Studies show that if the
opioids system necessities are not naturally supplied, people tend to present dysphoric humor, discomfort and
irritability, that seems to be reverted through manipulation of other people, alcohol and psychoactive
substances abuse, self-aggression (releasing endorphin), increase in the frequency of sexual relations,
impulsivity or experimentation of new sensations, observed in psychopathy. There are also speculations that
insufficient oxytocin can play an important role in the disorders pathogeny. Conclusion: Indeed, the
psychopathy constitutes a real challenge to the clinical understanding and to the patients management, owing
the multiple variables underlying the antisocial behavior and the emotional absence. In face of this complexity,
the knowledge of neurobiological bases open the doors not only to a more global comprehension of the
problem, but also to the developing of new treatments and assistance forms.
Financial Support: None.
Key words: psychopathy; behaviors; neurobiological bases

POMEGRANATE SEED OIL NANOEMULSIONS AS NANOCARRIER FOR


A NON-STEROIDAL ANTI-INFLAMMATORY DRUG: IMPROVEMENT OF
ANTINOCICEPTIVE EFFECT IN MICE
Luana Mota Ferreira1, Vernica Ferrari Cervi1, Marcel Henrique Marcondes Sari2,
Vanessa Angonesi Zborowski2, Cristina Wayne Nogueira2, Letcia Cruz1.
1

Programa de Ps-Graduao em Cincias Farmacuticas, Universidade Federal de Santa Maria.


Programa de Ps-Graduao em Bioqumica Toxicolgica, Centro de Cincias Naturais e Exatas,
Universidade Federal de Santa Maria.
2

Introduction: The use of vegetable oils has increased in the pharmaceutical field due to their biological
potential, which can improve the therapeutic effect of drugs. Pomegranate seed oil (PSO) has been attracted
interest due to its anti-inflammatory property. Nanoemulsions (NEs) present advantages including tailoring of
drug release, increase gastrointestinal distribution and protection of fragile drugs. Objective: This study aimed
to develop PSO based NEs loaded with ketoprofen (KP) to propose a new alternative for pain and inflammation
management. Methods: PSO NEs were prepared by the spontaneous emulsification method employing PSO
at 3.0 % and KP at 1 mg/mL. NEs were characterized by pH, droplet sizes, polydispersity, zeta potential, drug
content and encapsulation efficiency. To performed the in vivo experiments (Ethical Research Committee
#041/2014) male adult Swiss mice (2535 g) were randomly assigned into different groups. The
antinociceptive effect was evaluated by the acetic acid induced abdominal constrictions after intraperitoneal
injection (1.6%, 10 mL/Kg). The animals were placed individually in separate cages and the abdominal
constrictions were counted cumulatively over a period of 20 min. A model of inflammatory persistent pain
induced by the intraplantar injection of Complete Freuds Adjuvant (CFA) was also performed. The evaluation
of mechanical allodynia testing at the Von Frey Hair filaments was conducted. The free KP (FKP), dissolved
in a saline dispersion of Tween 80 1%, and NEs were administered by the intragastric route in a single
administration and in a constant volume of 10 mL/Kg. The dose- and time response curves were conducted to
each test. By statistical analyses, p values < 0.05 were considered statistically significant. Results: NEs
presented characteristics compatible with nanostructured systems for drug delivery. By acid acetic test FKP
(10 mg/Kg) triggered antinociceptive action at 0.5 h and remained significant up to 2 h. NEs started the effect
at 0.5 h and kept up to 12 h. The dose-response curve showed that 5, 1 and 0.5 mg/Kg FKP significantly
reduced the number of writhes whereas NEs showed effect at 5 and 1 mg/Kg. CFA test showed that the FKP
(10 mg/Kg, i.g.) reduced the mechanical allodynia at 0.5 h and remaining significant up to 6 h. NEs (10 mg/Kg,
i.g) reduced the mechanical response stimulus from 0.5 h up to 10 h. Dose-response curves showed that FKP
and NEs also presented antiallodynic effect at 5 mg/Kg. Conclusion: Nanoemulsions prolonged the KP
antinociceptive effect, demonstrating an improvement of its pharmacological activity.
Financial Support: CAPES.
Key words: pomegranate seed oil; nanoemulsions; antinociceptive activity.

INFLUENCE OF MAGNESIUM SUPPLEMENTATION ON THE


HALOPERIDOL-INDUCED COGNITIVE DEFICITS
Lusa Weber1, Maikel Kronbauer2, Vinicia G. Metz1, Marilise E. Brger1,2,3
1

Departamento de Fisiologia e Farmacologia, Universidade Federal de Santa Maria


Programa de Ps-Graduao em Farmacologia, Universidade Federal de Santa Maria Programa de PsGraduao em Farmacologia, Universidade Federal de Santa Maria.
3
Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
2

Introduction: Typical antipsychotics are the most prescribed worldwide drugs to treat mental disorders, whose
use is related to motor disturbances and worsening cognitive dysfunction in schizophrenic patients. Magnesium
(Mg) is the second most abundant intracellular cation in the body and plays important physiological functions
such as enzymes cofactor, synaptic plasticity and modulation of antioxidant defenses. Several studies have
shown that the increasing Mg level in the brain is important to regulation of synaptic plasticity and enhance
cognitive functions. Objective: The aim of this study was to investigate the influence of Mg supplementation
on cognitive deficits induced by haloperidol using the y-maze task. Methods: Twenty-four rats that were
designated to four experimental groups (n = 6) received haloperidol injection (H and MgH group 12
mg/Kg/mL) or vehicle (C and Mg groups), once a week for four weeks. Concomitantly, animals were orally
supplemented with magnesium aspartate (Mg and MgH groups 40 mg/Kg/mL) or deionized water (C and H
groups). Seven days after the last H administration, animals were submitted to y-maze behavioral paradigm
(for 5 min) for evaluations of sequence and total number of arms entered and percentage alternation score. All
behavioral measures were assessed using the ANY-MazeTM software. Results: Haloperidol administration
reduced both total arms entries and % of alternation. Although Mg supplementation did not alter the reduced
total arms entries it significantly exerted protection on the % of reduction of alternation induced by haloperidol.
Conclusion: These data confirm the deleterious effects of haloperidol treatment on memory functions,
suggesting that Mg supplementation could be used as supporting therapy in the antipsychotic treatment, thus
reducing these side effects.
Financial Support: CNPq, CAPES, FAPERGS.
Key words: magnesium supplementation; cognitive deficits; haloperidol.

(OMEPHSE)2-LOADED POLYMERIC NANOCAPSULES AS A NOVEL


APPROACH TO INFLAMMATORY PAIN TREATMENT: BEHAVIORAL
AND MOLECULAR EVIDENCE
Marcel H. M. Sari1, Vanessa A. Zborowiski1, Natlia da Silva Jardim1, Luana M.
Ferreira2, Letcia Cruz2, Cristina W. Nogueira1
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de


Santa Maria.
2
Programa de Ps-Graduao em Cincias Farmacuticas, Universidade Federal de Santa Maria.
Introduction: Inflammation is a complex and protective process but its excessive response could damage
healthy tissue and requires pharmacological intervention. Due to the several biological properties attributed to
organoselenium compounds, this class emerges as a source of new potential candidates to treat some disorders,
including the inflammatory process. However, some physicochemical limitations hinder more advanced
studies. The incorporation to nanostructured systems arises as a promising alternative to overcome such
restrictions and even improve biological effect of compounds. Objective: This study investigated the
protective effect exerted by p,p-methoxyl-diphenyl diselenide [(OMePhSe)2], incorporated into polymeric
nanocapsules in the behavioral and molecular alterations induced by an inflammatory pain model. Methods:
The (OMePhSe)2 nanocapsules suspension [NC (OMePhSe)2] was prepared through the interfacial deposition
of preformed polymer technique. Male Swiss mice (CEUA n 1446300315/2015) received a hindpaw
intraplantar injection of complete Freunds adjuvant (CFA) and 24 hours later they were treated through the
intragastric route with a single (OMePhSe)2 administration, in its free form (dissolved in canola oil) or NC
(OMePhSe)2, at a dose of 25 mg/kg. A time-response curve was conducted to evaluate the possible antiallodynic effect using the von Frey hair test. Celecoxib (30 mg/kg, single intragastric administration) was used
as positive control. Another set of animals were treated as described and at a time in which the anti-allodynic
effect of each treatment reached its peak, cerebral contra-lateral cortex samples were collected in order to
evaluate the immunocontent of some proteins through western blot. The results were evaluated by One-way
ANOVA followed by the NewmanKeuls' test. Results: Regardless its form, treatment with (OMePhSe)2 or
celecoxib reduced the allodynia caused by CFA injection. The free compound and celecoxib decreased the
allodynic response 0.5 h after treatment and their effects remained significant until 4 h later, with a peak in 1
and 3 h, respectively. The NC (OMePhSe)2 anti-allodynic effect started in 1 h and remained significant for up
to 7 h after its administration, with a peak in 4 h. The western blot analyses showed that the treatment with
(OMePhSe)2, free or NC, reduced the MAPK pathway activation (JNK; p38; ERK) as well as iNOS, COX-2,
NfB and interleukin 1 protein levels. Conclusion: The treatment with (OMePhSe)2 reduced the behavioral
and molecular alterations induced by CFA injection and its incorporation into nanocapsules prolonged its
effect.
Financial Support: UFSM, CNPq, CAPES, FAPERGS.
Key words: Organoselenium; Nanocapsules; Inflammation.

Buddleja thyrsoides LAM. CRUDE EXTRACT PRESENTS


ANTINOCICEPTIVE EFFECT IN A ARTHRITIC PAIN MODEL IN MICE
Maria Fernanda Pessano Fialho1, Indiara Brusco1, Sara Marchesan Oliveira1
1

Departamento de Bioqumica e Biologia Molecular, Universidade Federal de Santa Maria

Introduction: Rheumatoid arthritis is a chronic inflammatory disease characterized by joint pain which
reduces the life quality of affected individuals. Therapeutic tools used for the treatment of inflammatory pain
usually are associated with several undesirable effects. Thus, the search for most effective and safer drugs is
necessary. Buddleja thyrsoides Lam (EBBt), known as barbasco or cambara, is popularly used in several
disorders including antirheumatic, anti-inflammatory and analgesic activities. Objective: We investigated the
antinociceptive and anti-inflammatory effect of the oral treatment from the EBBt in a model of arthritic pain.
Methods: Were used male adult mice (25-35 g) (CEUA n1259221015/2016). For induction of arthritic pain
model, the animals received an intraplantar CFA injection (20 L, i.pl.) and after 48 hours they were orally
treated with EBBt (30-300 mg/kg) or vehicle (10 ml/kg, p.o.). The mechanical allodynia, thermal hyperalgesia
and edema were evaluated by filaments von Frey, Hargreaves test and digital micrometer, respectively. To
assess if EBBt reduces the leukocyte infiltration CFA-induced, was measured MPO activity (marker for
neutrophils), NAGase activity (marker for macrophages), and it was verified histological changes in the plantar
tissue. Moreover, it was also evaluated if EBBt cause adverse effects. Results: The EBBt oral treatment
reversed the mechanical allodynia and thermal hyperalgesia induced by CFA with maximum inhibition (Imax)
of 696% and 7815%, respectively. On the other hand, EBBt was unable to alter the paw edema. The EBBt
neither alter the MPO and NAGase activity nor inhibit the CFA-induced cellular infiltration. Finally, the oral
treatment was unable to cause adverse effects such as alter the body temperature, motor performance and
biochemical parameters or induced gastric lesions. Conclusion: These findings suggest that the treatment oral
of EBBt has antinociceptive but not anti-inflammatory effect in an arthritic pain model and could be used to
treat arthritic pain.
Financial Support: CNPq, CAPES and FIT BIT.
Key words: Barbasco; Cambara; nociception; pain; arthritis.

EFFECTS OF FUMONISIN B1 ON BEHAVIORAL TESTS IN MICE


Micheli Dassi1, Ana Claudia Monteiro Braga2, Naili Schiefelbein Souto1, rica Furlan
da Rosa2, Ana Flvia Furian1,2
1
2

Programa de Ps-Graduao em Cincia e Tecnologia em Alimentos, Universidade Federal de Santa Maria.


Programa de Ps-Graduao em Farmacologia, Universidade Federal de Santa Maria

Introduction: Fumonisins are mycotoxins mainly produced by Fusarium spp. mold, which constitutes a major
public health issue because of its worldwide distribution and diversity of toxic effects. While the liver and
kidney are considered the major target organs of fumonisin B1 (FB1) toxicity in several species, evidence
indicates that FB1 may be toxic to the brain. In addition, this mycotoxin is related to several damages to human
and animal health, among them neurodegenerative processes and development of depressive and anxiety
behavior. Objective: The aim of the present study was to evaluate the behavioral effects caused by FB1
exposure in mice. Methods: Male mice C57BL/6J (20-30g) received four administrations of FB1 (8 mg/kg,
i.p., 10 ml/kg, 1x/day) or its vehicle (0.9% NaCl e DMSO 1.6%, i.p., 10 ml/kg) and after the last administration
behavioral analyses were performed. We tested the animals on open field test, object recognition and nest
construction. The protocols were previously approved by the Ethics Commission on Animal Use of the Federal
University of Santa Maria under process number n 6775010416. Results: FB1 does not alter the number of
crossings, rearings and time spent in periphery, neither the recognition index observed in the object recognition
test. However, an increase in anxiety of the animals that received FB1 was observed in nest construction score.
Conclusion: These data suggest that repeated exposure to FB1 increases anxiety in animals without any
alterations on locomotor and exploratory activites.
Financial Support: CNPq, FAPERGS.
Key words: mycotoxin; behavior; exploratory activity; anxiety.

ANXIOLYTIC-LIKE EFFECTS OF TAURINE IN ZEBRAFISH SUBJECTED


TO THE LIGHT-DARK TEST
Nathana J. Mezzomo1, Vanessa A. Quadros2, Denis B. Rosemberg2
1

Programa de Ps-Graduao em Farmacologia, Universidade Federal de Santa Maria.


Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de
Santa Maria.
2

Introduction: Taurine (TAU) is an amino sulfonic acid with several functions in central nervous system.
Mounting evidence suggests that it acts in osmoregulation, neuromodulation and as an inhibitory
neurotransmitter. However, the effects of TAU on behavioral functions, especially on anxiety-related
parameters, are limited. The adult zebrafish is a suitable model organism to examine anxiety-like behaviors
since it presents neurotransmitter systems and behavioral functions evolutionary conserved. Anxiety in
zebrafish can be measured by different tasks, analyzing the habituation to novelty, as well as the response to
brightly lit environments. Objective: The aim of this study was to investigate whether acute TAU treatment
alters anxiety-like behavior in zebrafish using the novel tank and the light-dark tests. Methods: Fish were
individually treated with TAU (42, 150, and 400 mg/L) directly added the tank water for 1h and the behaviors
were further analyzed for 6 min in the novel tank or in the light-dark test. Control fish were handled in a similar
manner, but kept only in home tank water. All behavioral measures were assessed using the ANY-MazeTM
software and the following endpoints were analyzed: distance traveled, absolute turn angle, number of entries
and transitions to the upper half of the test tank (novel tank test), shuttling, time spent in the lit area and number
of risk assessment episodes (light-dark test). The protocols were previously approved by the Ethics
Commission on Animal Use of the Federal University of Santa Maria under process number 026/2014.
Results: Although TAU did not alter locomotor and vertical activities, all concentrations significantly
increased shuttling and time spent in lit compartment. Moreover, TAU 150 group showed a significant decrease
in the number of risk assessment episodes. Conclusion: These data suggest that TAU exerts an anxiolytic-like
effect in zebrafish and the comparative analysis of behavior using different tasks is an interesting strategy for
neuropsychiatric studies related to anxiety in this species.
Financial Support: CNPq, CAPES, FAPERGS.
Key words: anxiety; behavior; exploratory activity.

ANESTHETIC AND ANTIOXIDANT ACTIVITIES OF A


SESQUITERPENOID OBTAINED FROM Nectandra grandiflora ESSENTIAL
OIL
Patrcia Rodrigues1, Quelen I. Garlet2, Luana S. Balconi1, Luana da C. Pires2, Bernardo
Baldiserotto1, Berta M. Heinzmann1,2
1

Curso de Farmcia, Centro de Cincias da Sade/CCS, Universidade Federal de Santa Maria/UFSM, Santa
Maria, RS, Brazil.
2
Programa de Ps-Graduao em Farmacologia, CCS, UFSM, Santa Maria, RS.
Introduction: Essential oils (EO) and their isolated constituents are an alternative source of antioxidant,
sedative and anesthetic compounds (FIGUEIREDO et al., 2008). In vitro antioxidant assays, as the reducingantioxidant power of iron (FRAP), have been reported as good predictors of in vivo antioxidant potential
(AMORATI and VALGIMIGLI, 2015). Natural products can be used in aquaculture to reduce fish stress
during handling processes and to minimize damage resulting from oxidative stress, which impairs the animal
growing (CHAKRABORTY; HANCZ, 2011). Therefore, this work aims to evaluate the sedative/anesthetic
and antioxidant potential of a sesquiterpene (SF) isolated from Nectandra grandiflora EO. Methods: Silver
catfish juveniles (Rhamdia quelen) (7.99 0.41g; 8.62 0.31cm) were maintained in continuously aerated
250L tank and controlled temperature (21 -23C), fed once a day with commercial diet (28.0% crude protein)
and kept fasting for 24h before experiments. Compound SF was diluted in ethanol 1:10 and added to aquarium
containing 1L water at concentrations of 50, 100 and 200mg/L (GOMES et al. 2003). In vitro antioxidant
activity was assessed by FRAP test, according to Benzie and Strain (1996) with modifications. SF was
dissolved in ethanol at concentrations of 45, 10, 4,5; 1,0 and 0,45g/L. Compound SF (20L) or quercetin
(positive control) were pipetted into a 96 well plate and followed by the addition of FRAP reagent (180L).
The plate was incubated at 37C for 4 min and the antioxidant activity was measured at 593nm (Espectro
Thermo Plate TP-READER) in triplicate. Results: All SF concentrations induced light sedation (partial loss
of posture). Compound SF at 100mg/L induced total loss of posture (fish lie in the aquarium bottom) and at
200mg/L induced anesthesia stage (total loss of posture without reaction to contact stimuli. Antioxidant activity
was detected at 10 and 45M SF. Conclusion: The isolated compound SF induced sedation and long-term
anesthesia without adverse behavioral effects or mortality and showed moderate antioxidant activity in vitro.
Further studies should be conducted to determine the mechanism of action and to evaluate other biological
activities of this compound.
Financial Support: FAPERGS and CNPq

ENVIRONMENTAL ENRICHMENT ALTERS ADULT ZEBRAFISHS (Danio


rerio) LOCOMOTOR PROFILE
Thain Garbino dos Santos, Ben Hur Marins Mussulini, Diogo Losch de Oliveira.
Laboratory of Cell Neurochemistry, Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica,
Departamento de Bioqumica, Instituto de Cincias Bsicas da Sade, Universidade Federal do Rio Grande
do Sul, Porto Alegre, Brazil.
Introduction: Environmental enrichment offers social, sensory and cognitive stimuli for the animal and it has
been demonstrated that this condition increases neuronal proliferation and induces cognitive improvements in
different animal models. Objective: The main goal of this present study was investigated if there are
differences in the zebrafishs behavioral profile when animals are subjected to environmental enrichment for
one week. Material and Methods: Adult wild-type zebrafish of both sexes were kept in optimal conditions
for temperature, pH and light/dark cycle. Animals were divided in two groups: standard environment (SE
empty aquarium) and enriched environment (EE - aquarium had stones at bottom and three green artificial
plants). Animals remained during seven days in respective environments until behavioral tests. Each animal
was used for a single behavioral test. Open tank (OT), black/white and social interaction (SI) tests were
performed. Data were express as mean SEM and analyzed by unpaired t test or ANOVA two-way repeated
measures. P < 0.05 was considering statistically significant (n = 16-20). Results: In OT test, EE did not alter
total mobile time but reduced total distance travelled and distance travelled on the apparatus bottom as well as
had lower speed when compared with SE group. In SI test, there was no difference on total mobile time, EE
also reduced total distance travelled and speed besides did not have alteration in social preference. Lastly, in
black/white test, there were not differences between groups. Conclusions: OT and SI tests data indicate that
EE group has a slower locomotor profile comparing with SE group. Environmental enrichment protocol of one
week on OT test also demonstrated that EE moved similarly during all duration of the test at the apparatus
bottom while SE has a lot of locomotion in the starting and a lot of more at the end of test.
Financial support: CNPq, FAPERGS, PROPESQ-UFRGS.
Key words: zebrafish; environmental enrichment; behavior.

EFFECT OF ACUTE AND CHRONIC EXPOSURE OF THE ALARM


SUBSTANCE (AS) ON THE ACTIVITY OF CEREBRAL MONOAMINE
OXIDASE IN ZEBRAFISH
Vanessa A.de Quadros, Denis Broock Rosemberg1
1

Programa de Ps-Graduao em Cincias Biolgicas: Bioqumica Toxicolgica, Universidade Federal de


Santa Maria.
Introduction: The monoaminoxidase (MAO) is a flavoprotein that has great importance in the regulation of
some neurotransmitters such as serotonin, dopamine and noradrenaline, and it is known that in stressful
situations there may be neurochemical changes as well as regulatory enzymatic alterations of the central
nervous system like the MAO. In this sense, the alarm substance (AS) can be considered an important tool
since it has the capacity to induce fear / anxiety responses in fish and other animals. With this, zebrafish (Danio
rerio) is an experimental model widely used for conducting behavioral and neurochemical experiments. These
animals are considered non-mammalian vertebrate alternative models and are known for their small size, low
cost, easy maintenance, similarity with mammals, including humans, and for presenting significant responses
in the expression of brain MAO under the alteration of the co-specific ones caused by the AS. Objective:
Therefore, the aim of this study was to evaluate the possible behavioral and neurochemical alterations in two
strains of zebrafish (WT and leo) in the acute and chronic exposure alarm substance in aggressive context.
Methods: The monoaminoxidase (zMAO) activity was determined using two zebrafish brains were pooled
per sample, homogenized in 0.5 mL of a buffer solution containing 16.8 mM Na2HPO4 and 10.6 mM KH2PO4
isotonized with sucrose, pH 7.4, and further centrifuged at 900 g for 5 min. A 100 L aliquot of the supernatant,
was removed and mixed to 460 L of assay buffer (168 mM Na2HPO4 and 10.6 mM KH2PO4, isotonized with
KCl, pH 7.4). Next, the samples were preincubated at 37C for 5 min. The reaction was started after the
addition of kynuramine dihydrobromide at a saturating concentration of 110 M in a total volume of 700 L.
After 30 min, the reaction was terminated with 300 L of 10% trichloroacetic acid (TCA) and the tubes were
kept on ice. Samples were further centrifuged at 16.000 g for 5 min, and 800 L of supernatant was mixed to
1 mL of 1 M NaOH. The fluorescence intensity was measured spectrofluorimetrically. Results: Our results
demonstrated that the formation of product is linear in the range of 60 to 120 g in 30 min of reaction. Acute
exposure to AS significantly decrease brain MAOz activity in zebrafish WT and not difference in leo, whereas
no significant effects were observed after chronic exposure in both strains. Conclusion: Finally, we conclude
that the determination of MAOz activity may be an important tool in studies related to models of
aggressiveness in the neurochemical alterations in zebrafish.
Financial Support: CNPq, CAPES, FAPERGS.
Key words: Agonistic behavior; zebrafish; monoamine oxidase.

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