Search Results (208)

Background: Subclinical psychological symptoms—such as low mood, perceived stress, and poor sleep—affect a large portion of the population and can impair quality of life despite remaining below clinical thresholds. The gut–brain axis has emerged as a promising target for interventions that support emotional and psychological resilience. Probiotics and postbiotics are gaining attention for their potential to modulate mood and stress via microbiome-related mechanisms, but human evidence remains limited, particularly in non-clinical populations. Objectives: We aimed to assess the effects of a two-strain combination of live microorganisms alongside a two-strain combination of heat-treated inactivated microorganisms on outcomes associated with anxiety, mood, perceived stress, and quality of life in healthy adults experiencing mild stress. Methods: This study was conducted in two parts. In Part I, a randomized, double-blind, placebo-controlled study, 100 participants were randomized to receive either a blend of live microorganisms (Bifidobacterium longum CECT 7347 and Lactobacillus rhamnosus CECT 8361) or an identical placebo once daily for 12 weeks. In Part II, a pilot feasibility study, a subset of eight placebo non-responders from Part I received the heat-inactivated preparation of the same bacterial strains in a 6-week trial extension phase. For Parts I and II, the primary outcome was the change in the Hamilton Anxiety Rating Scale (HAM-A). Secondary outcomes included measures of mood (Beck Depression Inventory (BDI); Patient Health Questionnaire-9 (PHQ-9)), stress (state and trait anxiety inventory (STAI); Perceived Stress Scale (PSS)), sleep (Pittsburgh Sleep Quality Index (PSQI)), quality of life (36-item Short Form Survey (SF-36)), gastrointestinal symptoms (Gastrointestinal Symptom Rating Scale (GSRS)), salivary cortisol and microbiome modulation. Results: In Part I, there were no significant effects of the live blend on the HAM-A, indicating that the primary endpoint was not met. In addition, no significant effects were seen on the STAI or PSS scores when compared to the placebo. However, participants consuming the live blend trended toward a reduction in total PHQ-9 scores compared to placebo (p = 0.089), whilst preliminary exploratory analyses suggested an improvement in anhedonia (p = 0.045). Furthermore, there was a significant improvement in the vitality domain of the SF-36 compared to placebo (p = 0.017). On microbiome analysis, it was noted that consumption of the live blend was linked to the preservation of butyrate-producing bacteria, particularly members of the Pseudoflavonifractor genus and the Clostridium SGB6179 species. Furthermore, the abundance of B. longum species was found to be inversely associated with the total PSS Scores. In Part II, supplementation with the inactivated preparation resulted in significant within-group improvements for the vitality (p = 0.006) and social functioning (p = 0.010) domains of the SF-36 and improvements in PSS scores compared to baseline (p = 0.050). Conclusions: Supplementation with either the dual-strain live or inactivated formulations was associated with significant improvements in the vitality domain of the SF-36, whilst participants receiving the inactivated formulation demonstrated lower perceived stress and improved social functioning compared to baseline. Overall, the findings from this pilot study suggest that these two biotic consortia are well-tolerated and may be associated with improvements in measures of vitality in individuals with subclinical psychological symptoms. The subtle observations detected for stress and anhedonia suggest that further well-powered trials are needed to better characterize these findings, potentially in populations with greater baseline symptomatology. Full article

Purpose: The aims of this study were to evaluate the feasibility of time-restricted eating (TRE) in patients with Cushing’s disease (CD) and assess its effects on body weight and metabolic parameters. Methods: Twelve CD patients in remission with obesity were enrolled in a TRE program restricting food intake to 10:00–18:00. Anthropometric data, glycemic and lipid profiles, and circadian cortisol secretion were assessed at baseline and post-intervention. Serum cortisol levels were measured at multiple time points to evaluate diurnal patterns. Results: Nine patients completed the study. Over the 12-week period, participants showed a significant reduction in body weight, with median values decreasing from 93.8 kg [83.1–106.5] to 82.6 kg [76.9–100.3] (p = 0.011). Body mass index (BMI) also declined from 37.6 kg/m² [34.2–39.7] to 34.4 kg/m² [32.6–38.3] (p = 0.012). No statistically significant changes were observed in fasting glucose, HbA1c, or lipid parameters. Notably, 24 h urinary free cortisol levels significantly decreased (p = 0.01), and serum cortisol showed a downward trend at all measured time points, with the most pronounced reductions during mid-day and evening hours. No clinical or biochemical evidence of CD relapse was observed during the 12-month follow-up. Conclusions: Time-restricted eating is a feasible and well-tolerated dietary approach for patients with CD in remission, promoting weight loss and modest improvements in metabolic markers and cortisol rhythmicity. Full article

The use of olive by-products in livestock farming is a valuable resource, given their high levels of bioactive compounds with antioxidant and health-promoting properties. This preliminary study adopted an integrated approach to evaluate the influence of dietary Olea europaea L. polyphenols on animal welfare, physiological stress response, intestinal morphofunctional traits, and meat quality in Neroametà finishing pigs, a novel Casertana × Large White genetic line (Neroametà). Thirty pigs reared under extensive farming conditions were randomly allocated to two groups (n = 15): a control group fed a standard diet (C) and a treatment group (OL) supplemented with 300 mg/head/day of olive polyphenolic extract for 90 days. The study focused on the systemic correlation between host health and product quality. Meat composition, rheological properties, meat antioxidant activity, stress parameters, and fatty acid profiles of the longissimus lumborum and psoas major muscles were analyzed. Results showed that the OL diet significantly modulated the HPA axis, as evidenced by a marked reduction in plasma ACTH and cortisol levels, alongside improved antioxidant status. These physiological changes were positively associated with a trophic effect on the intestinal mucosa, characterized by increased villus height and a more favorable villus/crypt ratio. Regarding meat quality, the OL group exhibited superior oxidative stability, optimized pH decline, and an improved intramuscular fatty acid profile (increased MUFA and n-3 PUFA, reduced SFA). Despite the pilot scale of 30 animals, these findings provide a solid foundation for characterizing the Neroametà breed. In conclusion, Olea europaea L. polyphenols act as a multi-level modulator, enhancing physiological resilience and meat quality, offering a sustainable strategy for high-quality pork production in line with circular economy and One Health principles. Full article

Background/Objectives: Adaptogens are plant-derived substances that enhance the body’s nonspecific resistance to physical, chemical, biological, and psychological stressors by normalizing physiological functions. This article discusses the molecular mechanisms of action of seven key plant adaptogens—Rhodiola rosea, Schisandra chinensis, Withania somnifera, Eleutherococcus senticosus, Panax ginseng, Ocimum tenuiflorum, and Bacopa monnieri—in the context of chronic stress and lifestyle-related diseases. Methods: A review of the scientific literature is performed, including preclinical in vitro and in vivo studies, randomized placebo-controlled clinical trials, and studies employing network pharmacology analyses, molecular docking, and genomic techniques such as gene expression profiling. The interactions of active constituents with signaling pathways, molecular targets, and synergistic mechanisms were analyzed based on publications from the years 2010–2025. Results: Adaptogens exhibit pleiotropic activity: they regulate the HPA axis (Hypothalamic–Pituitary–Adrenal axis); induce Hsp70/Hsp16 expression; modulate SAPK/JNK, FOXO, and NF-κB pathways; and demonstrate antioxidant and mitoprotective effects. Specific mechanisms include: salidroside from R. rosea activating PI3K/Akt; schizandrin B from S. chinensis stimulating Hsp70; withanolides from W. somnifera inhibiting PDE4D; ginsenosides from P. ginseng suppressing FKBP51; and bacosides from B. monnieri enhancing acetylcholine synthesis. Clinical studies confirm reductions in cortisol levels (14–30%), decreased fatigue, and improved cognitive function without adverse effects. Conclusions: Understanding the molecular mechanisms of adaptogens supports their application in integrative medicine for the treatment of stress-related disorders, depression, anxiety, and neurodegenerative diseases. Further clinical studies are needed to optimize dosages and standardize extracts. Full article

Background: For decades, the ketogenic diet has been successfully used for the treatment of obesity, metabolic syndrome, and type 2 diabetes. The mechanisms through which it affects metabolism are not fully understood, but the hormonal changes that occur during ketogenic nutrition are likely to play an important role. Objectives: To investigate the effect of the ketogenic diet on various hormones associated with obesity and the accompanying metabolic disorders in childhood. Methods: One hundred children aged 8–18 years with obesity were enrolled. After baseline anthropometric, biochemical, and hormonal testing, they followed a 4-month “well-formulated ketogenic diet.” Fifty-eight of them successfully completed the study with follow-up assessments. Among them, 8 girls had polycystic ovary syndrome (PCOS) and 7 children had Hashimoto’s autoimmune thyroiditis. Results: At the end of the 4-month period, there was a significant decrease in basal insulinemia (p < 0.0001) and in mean morning cortisol levels (p = 0.04), as well as an increase in adiponectin levels (p = 0.04). All girls with PCOS experienced spontaneous menstrual cycles, accompanied by a reduction in testosterone levels. TSH levels showed no change for the whole group (p = 0.13), but there was a significant decrease in T3 (p < 0.0001) and a mild increase in T4 (p = 0.05). Among patients with Hashimoto’s thyroiditis, TSH levels were significantly higher at the end of the study. Conclusions: A short-term, well-formulated ketogenic diet in children with obesity is associated with hormonal changes that support weight loss and improve insulin sensitivity. The diet shows particularly beneficial effects in girls with PCOS and may be considered as part of a comprehensive therapeutic approach in these patients. Monitoring thyroid function during ketogenic nutrition is advisable in patients with hypothyroidism and thyroid disorders. Full article

Background: Parkinson’s disease (PD) is a progressive neurodegenerative disorder characterized not only by motor impairments but also by debilitating non-motor symptoms (NMS) such as anxiety, depression, fatigue, and sleep disturbances. These symptoms are often resistant to dopaminergic therapies and significantly impact patients’ quality of life. Whole-body cryostimulation (WBC) has emerged as a promising non-pharmacological intervention with potential effects on stress modulation and psychological well-being. Materials and Methods: In total, 14 patients with idiopathic PD underwent 10 WBC sessions (−110 °C for 2 min) over two weeks. Blood samples for cortisol and serotonin were collected before and after the first and last sessions. Patients completed standardized questionnaires evaluating anxiety (STAI-Y1, Y2), depression (BDI), fatigue (FSS), sleep quality (SCI), and daytime sleepiness (ESS) at baseline and after the final session. Results: Significant reductions in serum cortisol and improvements in serotonin levels were observed. Psychological assessments showed decreased anxiety and depression scores, with improvements in fatigue and sleepiness (p < 0.05 for most scales). Discussion and Conclusions: Repeated WBC sessions were safe and well-tolerated and were associated with biochemical and psychological improvements. These preliminary findings support WBC as a promising complementary intervention for alleviating NMS in PD. Further randomized controlled studies are warranted to confirm these results. Full article

Background/Objectives: Forest bathing (Shinrin-yoku) is a nature-based approach with potential preventive health relevance. This review summarizes evidence on its effects on immune function, stress physiology, and neuroprotective pathways. Methods: A narrative review of peer-reviewed studies was conducted using major scientific databases, including observational and interventional research assessing physiological or neurocognitive outcomes following forest exposure. Results: Forest bathing is associated with enhanced natural killer (NK) cell activity, modulation of inflammatory cytokine profiles, reductions in cortisol levels, and shifts toward parasympathetic autonomic dominance. Evidence also suggests a contributory role of tree-derived biogenic volatile organic compounds and phytoncides in immune and stress-regulatory effects. Emerging findings indicate potential benefits for cognitive restoration, emotional regulation, and neurotrophic signaling; however, substantial heterogeneity in study design, exposure characteristics, and outcome measures limits direct comparability and causal inference. Conclusions: Current evidence supports forest bathing as a promising, low-risk strategy for supporting immune resilience, stress regulation, and neurocognitive well-being within a preventive health framework. Preliminary findings also suggest potential benefits in chronic neurological conditions, supporting its neuroprotective role within multimodal neurorehabilitation strategies. Standardized intervention protocols, mechanistic biomarkers, and longitudinal studies are required to strengthen clinical relevance and guide evidence-based integration into public health and lifestyle medicine. Full article

This study investigates the effects of physical activity on serum cortisol levels and phagocytic capacity of the innate immune system in eight common bottlenose dolphins under human care. Analysis of 8 pairs (16 samples) revealed a significant increase in cortisol during periods of physical activity (mean increase of 1.27 µg/dL, 122% elevation), accompanied by decreased phagocytosis in granulocytes (92% reduction) and monocytes (52% reduction). Statistical analyses demonstrated consistent negative correlations between cortisol levels and phagocytic function, suggesting that physical activity influences hypothalamic–pituitary–adrenal axis activation and, consequently, innate immune system function. The male showed an attenuated cortisol response, but maintained monocyte sensitivity. These findings highlight the complex interplay between the neuroendocrine cortisol response and immune function in cetaceans, for the management and evaluation of animals under human care. A multi-method statistical framework incorporating Bayesian analysis, bootstrapping, and traditional approaches ensured robust inference despite limited sample size. Full article

Background/Objectives: Dental anxiety is a common clinical problem that negatively affects patient cooperation, treatment acceptance, and physiological stability during dental procedures. This randomized controlled clinical trial study aimed to evaluate the effectiveness of music-based interventions in reducing dental anxiety and stress responses during restorative dental treatment. The null hypothesis was that music exposure would not result in significant differences in anxiety levels or physiological stress parameters compared with standard dental care. Methods: Seventy-five patients with moderate to high pre-treatment dental anxiety (MDAS ≥10) were randomly assigned to three groups: classical music, Turkish music, and control (no music) (n = 25 per group). Anxiety levels were assessed using the Modified Dental Anxiety Scale (MDAS). Restorations were performed using a standardized adhesive protocol. Physiological parameters, including systolic and diastolic blood pressure (SBP, DBP), heart rate (HR), and oxygen saturation (SpO₂), as well as salivary cortisol and alpha-amylase levels, were measured before and after restorative treatment. Salivary cortisol and amylase levels were measured using a Human ELISA Kit. Statistical analysis was performed using paired t-tests and one-way ANOVA with Tukey’s post hoc test (p < 0.05). Results: Both music groups showed significant reductions in SBP, DBP, HR, cortisol, amylase, and MDAS scores compared to the control group (p < 0.05). Oxygen saturation increased significantly in the music groups, whereas it decreased significantly in the control group. There were no significant differences between classical and Turkish music regarding their anxiety-reducing effects. Conclusions: Music-based interventions effectively reduce dental anxiety and physiological stress during restorative dental procedures. This study is novel in simultaneously evaluating subjective anxiety scores and multiple physiological and biochemical stress markers in adult patients undergoing restorative treatment, supporting music as a simple and non-invasive adjunct in clinical dentistry. Full article

Aromatherapy, the therapeutic use of essential oils, is increasingly recognized as a complementary approach to women’s mental health, particularly during hormonally sensitive life stages such as menstruation, pregnancy, postpartum, and menopause. Concerns about the side effects of pharmacological treatments during these periods have driven interest in non-pharmacologic interventions. This narrative review synthesizes current clinical evidence on the efficacy of aromatherapy in alleviating psychological distress in women. A comprehensive literature review between 2000 and 2025 across PubMed, Scopus, and Web of Science databases identified 47 studies focusing on essential oils for anxiety, depression, or stress in female populations. The most substantial evidence supports the use of lavender, bergamot, rose, chamomile, clary sage, and ylang-ylang, with inhalation and massage as the most frequently studied delivery methods. Outcomes include reductions in cortisol, heart rate, and subjective stress, along with improvements in mood and emotional regulation. Aromatherapy demonstrates particular promise in postpartum and perimenopausal care. However, methodological heterogeneity and variability in oil composition limit generalizability. Despite these challenges, the evidence suggests that aromatherapy may serve as a safe, low-cost adjunct for managing mood disorders and stress in women, particularly when integrated into personalized, holistic care strategies. Full article

Background/Objectives: Chronic stress leads to sustained elevations in cortisol levels, which promote neuronal damage and impair memory. Prolonged stress also enhances proinflammatory signaling. Adaptogens are plant-derived compounds associated with the ability to increase the body’s resistance to stress, thereby improving mental and physical performance. To identify potential interventions capable of attenuating stress-related memory alterations, this study investigated a formulation combining the adaptogen Bacopa monnieri L. with phosphatidylserine and choline (BPC). Methods: An in vitro model of stress-related neuroinflammation was established by exposing BV2 microglial cells to corticotropin-releasing hormone (CRH, 100 nM). SH-SY5Y cells exposed to conditioned medium from CRH-stimulated BV2 cells or to iron(II) sulfate and L-ascorbic acid (Fe/Asc) were used as models of neurotoxicity. Results: BPC attenuated CRH-induced proinflammatory microglial morphology, as well as the reduction in cell viability and cell number. BPC treatment restored the levels of stress-related markers, including SIRT-1, Nrf-2, and phosphorylated JNK (p-JNK). Furthermore, BPC protected against neurotoxicity induced by CRH and Fe/Asc and promoted cholinergic activation by restoring basal acetylcholinesterase (AChE) levels. The combined BPC formulation showed superior efficacy compared with its individual components across all experimental assays. Conclusions: Collectively, these findings indicate that the BPC formulation developed in this study effectively attenuates stress-related neuroinflammation and neurotoxicity. BPC may represent a promising strategy to help limit the progression of early cognitive dysfunction under conditions of prolonged stress. Full article

Background/Objectives: Early childhood caries (ECC) and saliva have been studied across disparate domains, including microbiome, fluoride, immune, oxidative-stress, and neuroendocrine research. However, the ECC–saliva literature has not previously been mapped as a connected system using modern natural language processing (NLP). This study treats PubMed titles and abstracts as data to identify major themes, emerging topics, and candidate neuroimmune axes in ECC–saliva research. Methods: Using the NCBI E-utilities API, we retrieved 298 PubMed records (2000–2025) matching (“early childhood caries” [Title/Abstract]) AND saliva [Title/Abstract]. Text was cleaned with spaCy and embedded using a transformer encoder; BERTopic combined UMAP dimensionality reduction and HDBSCAN clustering to derive thematic topics. We summarised topics with class-based TF–IDF, constructed keyword co-occurrence networks, defined an internal topic-level Novelty Index (semantic distance plus temporal dispersion), and mapped high-novelty topics to gene ontology and Reactome pathways using g:Profiler. Prophet was used to model temporal trends and forecast topic-level publication trajectories. Finally, we generated a fully synthetic neuroimmune salivary dataset, based on realistic ranges from the literature, to illustrate how the identified axes could be operationalised in future ECC cohorts. Results: Seven coherent ECC–saliva topics were identified, including classical microbiome and fluoride domains as well as antioxidant/redox, proteomic, peptide immunity, and Candida–biofilm themes. High-novelty topics clustered around total antioxidant capacity, glutathione peroxidase, superoxide dismutase, and peptide-based host defence. Keyword networks and ontology enrichment highlighted “Detoxification of Reactive Oxygen Species”, “cellular oxidant detoxification”, and cytokine-mediated signalling as central processes. Temporal forecasting suggested plateauing growth for classical epidemiology and fluoride topics, with steeper projected increases for antioxidant and peptide-immunity themes. A co-mention heatmap revealed a literature-level Candida–cytokine–neuroendocrine triad (e.g., Candida albicans, IL-6/TNF, cortisol), which we propose as a testable neuro-immunometabolic hypothesis rather than a confirmed mechanism. Conclusions: AI-assisted topic modelling and network analysis provide a reproducible, bibliometric map of ECC–saliva research that highlights underexplored antioxidant/redox and neuroimmune salivary axes. The synthetic neuroimmune dataset and modelling pipeline are illustrative only, but together with the literature map, they offer a structured agenda for future ECC cohorts and mechanistic studies. Full article

Introduction: Massage therapy delivers structured mechanosensory input that can influence brain function, yet the central mechanisms and potential for neuroplastic change have not been synthesized across neuroimaging modalities. This mechanistic review integrates evidence from electroencephalography (EEG), functional MRI (fMRI), and functional near-infrared spectroscopy (fNIRS) to map how massage alters human brain activity acutely and over time and to identify signals of longitudinal adaptation. Materials and Methods: We conducted a scoping, mechanistic review informed by PRISMA/PRISMA-ScR principles. PubMed/MEDLINE, Cochrane Library, Google Scholar, and ResearchGate were queried for English-language human trials (January 1990–July 2025) that (1) delivered a practitioner-applied manual massage (e.g., Swedish, Thai, shiatsu, tuina, reflexology, myofascial techniques) and (2) measured brain activity with EEG, fMRI, or fNIRS pre/post or between groups. Non-manual stimulation, structural-only imaging, protocols, and non-English reports were excluded. Two reviewers independently screened and extracted study, intervention, and neuroimaging details; heterogeneity precluded meta-analysis, so results were narratively synthesized by modality and linked to putative mechanisms and longitudinal effects. Results: Forty-seven studies met the criteria: 30 EEG, 12 fMRI, and 5 fNIRS. Results: Regarding EEG, massage commonly increased alpha across single sessions with reductions in beta/gamma, alongside pressure-dependent autonomic shifts; moderate pressure favored a parasympathetic/relaxation profile. Connectivity effects were state- and modality-specific (e.g., reduced inter-occipital alpha coherence after facial massage, preserved or reorganized coupling with hands-on vs. mechanical delivery). Frontal alpha asymmetry frequently shifted leftward (approach/positive affect). Pain cohorts showed decreased cortical entropy and a shift toward slower rhythms, which tracked analgesia. Somatotopy emerged during unilateral treatments (contralateral central beta suppression). Adjuncts (e.g., binaural beats) enhanced anti-fatigue indices. Longitudinally, repeated programs showed attenuation of acute EEG/cortisol responses yet improvements in stress and performance; in one program, BDNF increased across weeks. In preterm infants, twice-daily massage accelerated EEG maturation (higher alpha/beta, lower delta) in a dose-responsive fashion; the EEG background was more continuous. In fMRI studies, in-scanner touch and reflexology engaged the insula, anterior cingulate, striatum, and periaqueductal gray; somatotopic specificity was observed for mapped foot areas. Resting-state studies in chronic pain reported normalization of regional homogeneity and/or connectivity within default-mode and salience/interoceptive networks after multi-session tuina or osteopathic interventions, paralleling symptom improvement; some task-based effects persisted at delayed follow-up. fNIRS studies generally showed increased prefrontal oxygenation during/after massage; in motor-impaired cohorts, acupressure/massage enhanced lateralized sensorimotor activation, consistent with use-dependent plasticity. Some reports paired hemodynamic changes with oxytocin and autonomic markers. Conclusions: Across modalities, massage reliably modulates central activity acutely and shows convergent signals of neuroplastic adaptation with repeated dosing and in developmental windows. Evidence supports (i) rapid induction of relaxed/analgesic states (alpha increases, network rebalancing) and (ii) longer-horizon changes—network normalization in chronic pain, EEG maturation in preterm infants, and neurotrophic up-shifts—consistent with trait-level recalibration of stress, interoception, and pain circuits. These findings justify integrating massage into rehabilitation, pain management, mental health, and neonatal care and motivate larger, standardized, multimodal longitudinal trials to define dose–response relationships, durability, and mechanistic mediators (e.g., connectivity targets, neuropeptides). Full article

Gastrointestinal nematodes (GIN) represent a major constraint to sheep production, and sustainable alternatives to routine anthelmintic use are increasingly required. This study compared two parasite control strategies in Zerasca sheep: routine albendazole treatment administered twice yearly (T) and a long-term non-chemical approach based on rotational grazing (relocation to a new pasture when grass height fell below 10 cm) combined with quarterly veterinary monitoring (NT). Twenty-four adult ewes (n = 12 per group) were monitored over an eight-month period. Mean faecal egg counts (EPG) differed significantly between groups (T: 210 ± 78; NT: 529 ± 89; p = 0.0007). In group T, EPG values decreased markedly 7 days after treatment but increased again by 150 days, resulting in no persistent differences between groups over time. Despite higher parasite burdens, NT sheep showed significantly higher body condition scores compared with treated animals (3.00 ± 0.61 vs. 2.51 ± 0.53; p = 0.0014). Haematological parameters were largely comparable between groups, although mild reductions in RBC, HGB, and HCT were observed in both. Treated sheep exhibited higher AST activity (p < 0.0001) and transient increases in ALT and BUN following treatment. Hair cortisol concentrations did not differ significantly between groups. Overall, these findings suggest that a non-chemical parasite management strategy, when combined with controlled grazing and veterinary monitoring, may maintain acceptable parasite levels while supporting body condition and stable welfare indicators, potentially reducing reliance on routine anthelmintic treatments. Full article

The most used technique to assess cortisol in zebrafish is trunk sampling, a terminal procedure. Extracting cortisol non-terminally in adult zebrafish remains challenging, limiting longitudinal studies, and the reduction of the number of zebrafish used in research. This study explored non-terminal methods for cortisol measurement in adult zebrafish under acute and chronic stress, focusing on housing water and skin mucus as alternatives to terminal trunk sampling. Oxidative stress markers (cerebral and hepatic) were also assessed to confirm stress responses. In experiment A, zebrafish were exposed to no stress, acute stress (AS), or chronic stress for 14 days (CS14) to evaluate skin mucus and trunk cortisol as biomarkers. In experiment B, in addition to CS14, a 7-day unpredictable chronic stress protocol (CS7) was tested to discard stress habituation. Results showed significant effects on cerebral oxidative stress: AS increased ROS and AChE activity, CS7 reduced GPx and AChE, and CS14 raised GPx in experiment A, while it increased protein carbonyls and decreased ATPase levels in experiment B. Trunk and skin mucus cortisol increased following AS. Under chronic stress, trunk and skin mucus cortisol levels were not significantly altered, but water cortisol increased at CS7. In conclusion, skin mucus and trunk cortisol levels are reliable biomarkers for acute stress, while water cortisol holds promise for chronic stress. Full article