Ethanol, also known as ethyl alcohol (EtOH), is a psychoactive ingredient present in various alcoholic beverages (Ngui et al., 2022). This psychoactive substance has properties that can cause chemical addiction and may cause a chronic and multifactorial disease known as alcohol use disorder (AUD) (OMS, 2018).

The chronic consumption of alcoholic beverages affects the metabolism of carbohydrates (by inhibiting gluconeogenesis), proteins (by increasing nitrogen excretion), and lipids, by increasing the esterification of fatty acids. As for micronutrients, it causes malabsorption, increased urinary excretion, and interferes with intestinal permeability, affecting the metabolism of vitamins such as thiamine, folate, retinol, ascorbic acid, and zinc, among others (Kamran et al., 2020).

Worldwide, the harmful use of alcohol results in three million deaths per year, in addition to having a cause-effect relationship with mental health disorders (CISA, 2022; OMS, 2018). Alcohol addiction is characterized by behavioral, cognitive, and physiological phenomena that are strongly associated with the desire to drink alcohol and difficulty controlling its consumption (Soyka et al., 2017).

Clinical studies have shown that alcohol withdrawal has high comorbidity with anxiety disorders. Chronic alcohol intake can result in addiction and increased stress and anxiety during abstinence periods (Campêlo et al., 2020). Therefore, AUD is often associated with psychiatric comorbidities, such as depression and anxiety disorder, which can be classified as the triggering factor or a coexistence framework, in which one can reinforce the other, when not treated correctly, requiring careful pharmacotherapy (Baltar et al., 2019).

The most recommended first-line pharmacological treatment for alcohol withdrawal syndrome is benzodiazepines (Teixeira, 2022). Despite being relatively safe, benzodiazepines have adverse effects that tend to resolve during their use, especially sleep-associated disorders, in addition to lethargy, sedation, and inability to perform daily activities (Mantovani and Quagliato, 2019).

The neuronal expression of neurotransmitters such as alpha 1 subunit-containing GABAA receptors is decreased during chronic alcohol consumption, and upon cessation of alcohol consumption (Lingford-Hughes and Nutt, 2003). This decrease in receptor levels causes a hypodopaminergic and hypoopioidergic environment in the reward system circuits, manifesting as withdrawal symptoms (Jukic et al., 2011).

The relationship between alcohol withdrawal and anxiety symptoms has not yet been adequately addressed, but alcohol withdrawal symptoms have been associated with the downregulation of type A γ-aminobutyric acid (GABAA) receptors and/or decreased GABAergic transmission (Sanna et al., 2003; Cagetti 2003; Melón et al., 2018). In this context, herbal medicines, and combined nutritional supplements are gaining attention for the development of new therapeutically active medications, with greater efficacy, lower toxicity, and fewer side effects than benzodiazepines (Gupta and Sharma 2019; Mantovani and Quagliato, 2019). The studies by Gupta and Sharma (2019), for example, detected that Bacopa monnieri, a medicinal plant from India, mitigates anxiety-like behavior induced by alcohol withdrawal, regulating the biochemical expression and expression of the Gabra1, Gabra4 Gabra5 genes of the GABAA receptor signaling pathway in rats.

Dietary supplements have gained worldwide prominence for their neuroprotective potential (Jukic et al., 2011; Li et al., 2021; Molin et al., 2019; Nitzan et al., 2022). For example, Jukic et al. (2011) tested the effects of dietary supplements containing D-phenylalanine, a peptidase inhibitor of opioid inactivation, and L-amino acids as a nutritional source for dopamine synthesis – to alleviate the symptoms of alcohol withdrawal, in 20 patients who suffered from alcohol dependence and started detoxification therapy. The researchers demonstrated that the administration of these dietary supplements reduced ethanol withdrawal symptoms.

Blue Calm® (BC) is a dietary supplement indicated to aid restorative sleep, aiming at relaxation and neuromuscular and energetic metabolic support. BC has traces of medicinal plant extracts as its major chemical constituents, namely myo-inositol, magnesium bisglycinate, taurine, and L-tryptophan, with potential effects on mental health, due to the improvement in the quality of sleep and relaxation (VIDA, 2023). These chemical constituents are associated with mental health disorders. Myo-inositol is involved in signaling pathways concerning different stimuli, such as the menstrual cycle hormones in women suffering from Polycystic Ovary Syndrome (PCOS) and, may positively influence psychological conditions related to depressive and anxious moods (Cantelmi et al., 2021). Bipolar disorder patients with anxiety symptoms have increased levels of inositol metabolism in the right prefrontal white matter (Chen et al., 2021). Magnesium (Mah and Pitre, 2021) and tryptophan have shown promising results in terms of improving sleep quality (Friedman, 2018; Binks et al., 2020; Bhat et al., 2021).

Because of the side effects of drugs such as benzodiazepines and the need for their use in the treatment of alcohol withdrawal symptoms, it is necessary to search for new substances that can have such benefits while not being associated with adverse effects. Thus, this study evaluated the potential of BC supplement in the treatment of alcohol withdrawal-induced anxiety in adul zebrafish (aZF).