Dietary Euterpe oleracea Mart. attenuates seizures and damage to lipids in the brain of Colossoma macropomum.

The Amazonian açai fruit (Euterpe oleracea) has shown promising anticonvulsant properties, comparable to those of diazepam (BDZ) in in vivo models submitted to pentylenetetrazole (PTZ). PTZ is a classic convulsant agent used in studies for the purpose of screening anticonvulsants and investigating the mechanisms of epilepsy. Herein, we aimed to determine, for the first time, the effect of dietary administration of lyophilized E. oleracea (LEO) on PTZ-induced seizures, using juvenile Colossoma macropomum fish (9.1 ± 1.5 g) as a model. A control diet (0.00% LEO) and two levels of LEO inclusion were established: 5.00% and 10.0% LEO (w/w). Fish were divided into five groups (n = 5): control (0.9% physiological solution; i.p.), PTZ (PTZ 150 mg kg-1; i.p.), PTZ LEO 5.00%, PTZ LEO 10.0%, and BDZ-PTZ (BDZ: diazepam 10 mg kg-1; i.p.). In addition to the electroencephalography (EEG), the lipid peroxidation (TBARS) was quantified in the brain, along with the characterization of behavioral responses. Fish receiving PTZ showed intense action potential bursts (APB), which overlapped with a hyperactive behavior. In PTZ LEO 5.00% and 10.0% groups, convulsive behavior was significantly reduced compared to the PTZ group. Fish fed 5.00% or 10.0% LEO and exposed to PTZ showed less excitability and lower mean amplitude in tracings. The inclusion of 10.0% LEO in the diet prevented the increase in mean amplitude of the EEG waves by 80%, without significant differences to the quantified mean amplitude of the BDZ-PTZ group. TBARS concentration was reduced by 60% in the brain of fish fed 10.0% LEO-enriched diets relative to the PTZ-administered group. The results of this study demonstrated the anticonvulsant and protective roles of LEO to the brain, and the dietary inclusion of LEO seems to be promising for the formulation of functional diets. Results of this study may boost the interest on the anti-seizurogenic properties of Euterpe oleracea, including the development of new approaches for the prevention of seizures in humans and animals with low epileptic threshold.

Eugenol induces body immobilization yet evoking an increased neuronal excitability in fish during short-term baths.

Several studies have suggested eugenol as a suitable anaesthetic for fish. However, it has also been regarded as a toxic and aversive substance to several aquatic organisms, including fish. This study sought to assess the eugenol-induced behavioural alterations and its seizurogenic potential to fish. Moreover, a distinctive methodology for an in vivo evaluation of the brain activity was also presented. Prior to the evaluation of eugenol-induced responses, fish were exposed to pentylenetetrazole (PTZ), to characterize any seizure-like patterns. Antagonizing responses to PTZ were assessed in fish receiving diazepam (BDZ) and subsequently exposed to PTZ. Tambaqui fish juveniles, Colossoma macropomum (15.8 ± 2.8 g) were used as models and assayed as follows: (i) fish exposed to PTZ (15 mM) and (ii) fish receiving a dose of BDZ (10 mg Kg-1) and later exposed to PTZ (15 mM) (BDZ-PTZ group). Thereafter, fish were evaluated throughout (iii) eugenol exposure at 65 µL L-1 (ethanolic solution) and recovery. Control fish and a vehicle control group (ethanol at 585 µL L-1) were also established. PTZ baths elicited body immobilization preceded by hyperactivity in a stereotyped seizure-like behaviour with increased EEG wave amplitude and frequency. PTZ effects in the brain were attenuated by a pre-administration of BDZ. Upon eugenol exposure, tambaqui had an intense neuronal excitability, showing a clonus-like seizure behaviour, also corroborated by the EEG patterns, which were consistent with a seizure-like response. Responses of eugenol-exposed fish resembled those of the PZT-exposed animals, with epileptiform discharges. EMG was in line with the EEG modulation, showing increased tracing oscillations and higher mean amplitudes in PTZ-exposed fish whereas in BDZ-PTZ group muscle contraction was less frequent and powerful. Fish exposed to eugenol showed initially some muscle activity followed by a loss of muscle tonus over time. In summary, our results showed that upon eugenol exposure, although a time-dependent body immobilization was attained, fish presented an intense neuronal excitability comparable to that evoked by PTZ. Eugenol failed to promote depression of the CNS and therefore may be not suitable to be used for general anaesthesia of C. macropomum. As eugenol could be implicated in seizurogenesis and be potentially toxic to the fish brain, protocols suggesting the broad use of eugenol for short-term anaesthesia or euthanasia of fish should be carefully revised, as it raises important concerns in terms of ethics and fish welfare.

Cunaniol-elicited seizures: Behavior characterization and electroencephalographic analyses.

This study aimed at describing the characteristics and properties of seizures induced by cunaniol, a polyacetylenic alcohol isolated from the Clibadium genus, which is ubiquitous in the Amazon biodiversity and its potential use as a convulsant model. Wistar rat behavior was assessed upon cunaniol administration and animals were evaluated for neural activity through electroencephalographic records whereby epidural electrodes were positioned over the motor cortex under cunaniol-elicited seizures and seizure's control using three anticonvulsant agents, namely phenytoin, phenobarbital and diazepam. Cunaniol-induced seizures displayed a cyclic development of electrocorticographic seizures, presenting interictal-like spike and ictal period, which correlates to the behavioral observations and is in line with acute seizures induced by pentylenetetrazole. Cunaniol-elicited seizures were intractable by phenytoin treatment and controlled under the GABAergic activities of phenobarbital and diazepam. The results indicate that the cunaniol-induced changes show characteristics of seizure activity, making this plant compound a suitable animal convulsant model for seizure-related studies that could be used to assist in the development of novel anticonvulsant agents.