The current study contributes to fill the knowledge gap on the neurotoxicity of inorganic mercury (iHg) in fish through the implementation of a combined evaluation of brain morphometric alterations (volume and total number of neurons plus glial cells in specific regions of the brain) and swimming behavior (endpoints related with the motor activity and mood/anxiety-like status). White seabream (Diplodus sargus) was exposed to realistic levels of iHg in water (2μgL(-1)) during 7 (E7) and 14 days (E14). After that, fish were allowed to recover for 28 days (PE28) in order to evaluate brain regeneration and reversibility of behavioral syndromes. A significant reduction in the number of cells in hypothalamus, optic tectum and cerebellum was found at E7, accompanied by relevant changes on swimming behavior. Moreover, the decrease in the number of neurons and glia in the molecular layer of the cerebellum was followed by a contraction of its volume. This is the first time that a deficit on the number of cells is reported in fish brain after iHg exposure. Interestingly, a recovery of hypothalamus and cerebellum occurred at E14, as evidenced by the identical number of cells found in exposed and control fish, and volume of cerebellum, which might be associated with an adaptive phenomenon. After 28 days post-exposure, the optic tectum continued to show a decrease in the number of cells, pointing out a higher vulnerability of this region. These morphometric alterations coincided with numerous changes on swimming behavior, related both with fish motor function and mood/anxiety-like status. Overall, current data pointed out the iHg potential to induce brain morphometric alterations, emphasizing a long-lasting neurobehavioral hazard.

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The current study aims to shed light on the neurotoxicity of MeHg in fish (white seabream - Diplodus sargus) by the combined assessment of: (i) MeHg toxicokinetics in the brain, (ii) brain morphometry (volume and number of neurons plus glial cells in specific brain regions) and (iii) fish swimming behavior (endpoints associated with the motor performance and the fear/anxiety-like status). Fish were surveyed for all the components after 7 (E7) and 14 (E14) days of dietary exposure to MeHg (8.7μgg), as well as after a post-exposure period of 28days (PE28). MeHg was accumulated in the brain of D. sargus after a short time (E7) and reached a maximum at the end of the exposure period (E14), suggesting an efficient transport of this toxicant into fish brain. Divalent inorganic Hg was also detected in fish brain along the experiment (indicating demethylation reactions), although levels were 100-200 times lower than MeHg, which pinpoints the organic counterpart as the great liable for the recorded effects. In this regard, a decreased number of cells in medial pallium and optic tectum, as well as an increased hypothalamic volume, occurred at E7. Such morphometric alterations were followed by an impairment of fish motor condition as evidenced by a decrease in the total swimming time, while the fear/anxiety-like status was not altered. Moreover, at E14 fish swam a greater distance, although no morphometric alterations were found in any of the brain areas, probably due to compensatory mechanisms. Additionally, although MeHg decreased almost two-fold in the brain during post-exposure, the levels were still high and led to a loss of cells in the optic tectum at PE28. This is an interesting result that highlights the optic tectum as particularly vulnerable to MeHg exposure in fish. Despite the morphometric alterations reported in the optic tectum at PE28, no significant changes were found in fish behavior. Globally, the effects of MeHg followed a multiphasic profile, where homeostatic mechanisms prevented circumstantially morphometric alterations in the brain and behavioral shifts. Although it has become clear the complexity of matching brain morphometric changes and behavioral shifts, motor-related alterations induced by MeHg seem to depend on a combination of disruptions in different brain regions.

There are several aspects of inorganic mercury (iHg) toxicokinetics in fish that remain undeveloped despite its environmental ubiquity, bioaccumulation capacity and toxicity. Thus, this study presents new information on the uptake, distribution and accumulation of iHg following water contamination by adopting a novel set of body compartments (gills, eye wall, lens, blood, liver, brain and bile) of the white sea bream (Diplodus sargus) over 14 days of exposure. Realistic levels of iHg in water (2 μg L(-1)) were adopted in order to engender reliable conclusions in the assessment of fish health. A depuration phase of 28 days was also considered with the purpose of clarifying iHg elimination. It was found that iHg was accumulated faster in the gills (within 1 day), which also had the highest accumulated levels among all the target tissues/organs. Moreover, iHg increased gradually with exposure time in all the tissues/organs, except for the lens that showed relatively unaltered levels throughout the experiment. After 14 days of exposure, lower values of Hg were recorded in the brain/eye wall compared to the liver, which is probably related with the presence of blood-organ protection barriers, which limit iHg influx. iHg reached the brain earlier than the eye wall (3 and 7 days, respectively) and, hence, higher accumulated levels were recorded in the former. A depuration period of 28 days did not allow the total elimination of iHg in any of the tissues/organs. Despite this, iHg was substantially eliminated in the gills, blood and liver, whereas the brain and eye wall were not able to eliminate iHg within this timeframe. The brain and eye wall are more "refractory" structures with regard to iHg elimination, and this could represent a risk for wild fish populations.

The effect of two anesthetics, 2‐phenoxyethanol (2‐PE) and clove oil (COil) were studied in meagre, Argyrosomus regius. This study aimed to determine the adequate dose of anesthesia for different aquaculture procedures, to assess a sedative or stressor effect of low anesthetic concentrations, and to assess the effect of low anesthetic doses at high fish densities for longer periods of time on fish survival and behavior. Anesthetics were tested at different concentrations: 2‐PE at 100, 250, 400, 550, and 700 mg/L; COil at 25, 40, 55, 70, and 85 mg/L. Meagre became anesthetized within 10 min when using concentrations above 250 mg/L for 2‐PE and above 40 mg/L for COil. Deep anesthesia was achieved at 700 and 85 mg/L, for 2‐PE and COil, respectively. The fish did not react when blood was collected at these concentrations. Plasma cortisol and glucose levels were similar between sedated meagre with both 2‐PE – 100 mg/L and COil – 10 mg/L, and the control (not sedated; P > 0.05). This suggests that low concentrations of these anesthetics induce similar stress response as handling without anesthesia during routine activities. No mortality was registered when meagre was maintained at high densities with low concentrations of each anesthetic for 2 h.