Ana Cristina Varrone Giacomini

Diazepam and Fluoxetine Decrease the Stress Response in Zebrafish

The presence of pharmaceutical products in the aquatic environment has been reported in several studies. However, the impact of these drugs on living organisms is still uncharacterized. Here, we investigated the effects of acute exposure to either diazepam or fluoxetine on the stress response in Danio rerio. We showed that diazepam and fluoxetine inhibited the stress axis in zebrafish. Intermediate concentrations of diazepam suppressed the stress response as measured by cortisol levels, whereas fluoxetine inhibited cortisol increase at concentrations similar to those found in the environment. These data suggest that the presence of psychoactive drugs in aquatic ecosystems could cause neuroendocrine dysfunction in fish.

Fluoxetine and diazepam acutely modulate stress induced-behavior.

Drug residue contamination in aquatic ecosystems has been studied extensively, but the behavioral effects exerted by the presence of these drugs are not well known. Here, we investigated the effects of acute stress on anxiety, memory, social interaction, and aggressiveness in zebrafish exposed to fluoxetine and diazepam at concentrations that disrupt the hypothalamic-pituitary-interrenal (HPI) axis. Stress increased the locomotor activity and time spent in the bottom area of the tank (novel tank). Fluoxetine and diazepam prevented these behaviors. We also observed that stress and fluoxetine and diazepam exposures decreased social interaction. Stress also increased aggressive behavior, which was not reversed by fluoxetine or diazepam. These data suggest that the presence of these drugs in aquatic ecosystems causes significant behavioral alterations in fish.

Acute exposure to waterborne psychoactive drugs attract zebrafish.

Psychotropic medications are widely used, and their prescription has increased worldwide, consequently increasing their presence in aquatic environments. Therefore, aquatic organisms can be exposed to psychotropic drugs that may be potentially dangerous, raising the question of whether these drugs are attractive or aversive to fish. To answer this question, adult zebrafish were tested in a chamber that allows the fish to escape or seek a lane of contaminated water. These attraction and aversion paradigms were evaluated by exposing the zebrafish to the presence of acute contamination with these compounds. The zebrafish were attracted by certain concentrations of diazepam, fluoxetine, risperidone and buspirone, which were most likely detected by olfaction, because this behavior was absent in anosmic fish. These findings suggest that despite their deleterious effects, certain psychoactive drugs attract fish.

My stress, our stress: blunted cortisol response to stress in isolated housed zebrafish.

Here, we show that individually housed zebrafish presented a reduced cortisol response to an acute stressor (persecution with a pen net for 120 s) compared to zebrafish housed in groups of 10. We hypothesized that the cortisol response to stress was reduced in individually housed zebrafish because they depend solely on their own perceptions of the stressor, whereas among grouped zebrafish, the stress response might be augmented by chemical and/or behavioral cues from the other members of the shoal. This hypothesis was based on previous described chemical communication of stress in fish as well on individual variation in stressor perception and potential individual differences in fish personality.

The smell of "anxiety": Behavioral modulation by experimental anosmia in zebrafish.

Olfaction is strongly involved in the regulation of fish behavior, including reproductive, defensive, social and migration behaviors. In fish, anosmia (the lack of olfaction) can be induced experimentally, impairing their ability to respond to various olfactory stimuli. Here, we examine the effects of experimental lidocaine-induced anosmia on anxiety-like behavior and whole-body cortisol levels in adult zebrafish (Danio rerio). We show that experimentally-induced anosmia reduces anxiolytic-like behavioral effects of fluoxetine and seems to interact with anxiogenic effect of stress also paralleling cortisol responses in zebrafish. These findings provide first experimental evidence that temporary anosmia modulates anxiety-like behaviors and physiology in adult zebrafish.

Effects of waterborne fluoxetine on stress response and osmoregulation in zebrafish.

The presence of fluoxetine in aquatic environments has been reported for decades. Here, we investigate the effects of exposure to fluoxetine on the stress response and osmoregulation in zebrafish. We show that stress response alters osmoregulation and that fluoxetine inhibits these stress-related changes in osmoregulation. The results suggest that the presence of fluoxetine in aquatic ecosystems can cause changes in response to stress and osmoregulation in fish.

Environmental and Pharmacological Manipulations Blunt the Stress Response of Zebrafish in a Similar Manner

Here we provide evidence that both pharmacological and environmental manipulations similarly blunt the cortisol release in response to an acute stressor in adult zebrafish. Different groups of fish were maintained isolated or group-housed in barren or enriched tanks, and then exposed or not to diazepam or fluoxetine. Acute stress increased cortisol levels in group-housed zebrafish maintained in barren environment. Single-housed zebrafish displayed a blunted cortisol response to stress. Environmental enrichment also blunted the stress response and this was observed in both isolated and group-housed fish. The same blunting effect was observed in zebrafish exposed to diazepam or fluoxetine. We highlighted environmental enrichment as an alternative and/or complimentary therapeutic for reducing stress and as a promoter of animal welfare.

Waterborne psychoactive drugs impair the initial development of Zebrafish

The contamination of rivers and other natural water bodies, including underground waters, is a current reality. Human occupation and some economic activities generate a wide range of contaminated effluents that reach these water resources, including psychotropic drug residues. Here we show that fluoxetine, diazepam and risperidone affected the initial development of zebrafish. All drugs increased mortality rate and heart frequency and decreased larvae length. In addition, risperidone and fluoxetine decreased egg hatching. The overall results points to a strong potential of these drugs to cause a negative impact on zebrafish initial development and, since the larvae viability was reduced, promote adverse effects at the population level. We hypothesized that eggs and larvae absorbed the drugs that exert its effects in the central nervous system. These effects on early development may have significant environmental implications.

Waterborne aripiprazole blunts the stress response in zebrafish

Here we provide, at least to our knowledge, the first evidence that aripiprazole (APPZ) in the water blunts the stress response of exposed fish in a concentration ten times lower than the concentration detected in the environment. Although the mechanism of APPZ in the neuroendocrine axis is not yet determined, our results highlight that the presence of APPZ residues in the environment may interfere with the stress responses in fish. Since an adequate stress response is crucial to restore fish homeostasis after stressors, fish with impaired stress response may have trouble to cope with natural and/or imposed stressors with consequences to their welfare and survival.

Evaluating "anxiety" and social behavior in jundiá (Rhamdia quelen)

Jundiá (Rhamdia quelen) is a suitable species for aquaculture in regions of temperate or subtropical climate. This species has received great attention regarding several aspects of physiology as well as an organism to study the impact of environmental contaminations. However, experiments using validated and objective tests to evaluate the jundiá behavior are scarce. The effects of acute stress have been studied in other fish species, such as zebrafish (Danio rerio), however, the effects in jundiá are lacking. Thus, we evaluated the effects of acute stress (net chasing) on anxiety-like and social behavior in jundiá. For these purpose, all behavioral analyses were carried out using automated tracking software. We showed that the acute stress protocol increased cortisol levels and induced anxiogenic-like behavior in the novel tank test, and decreased social behavior in jundiá. The antidepressant fluoxetine was able to prevent the effects of acute stress on social behavior. Here we show a behavioral evaluation of Rhamdia quelen using consolidated tests and computerized analysis, which allows more measurable, reliable and comparable results.