The Effect of Probiotic Supplementation on Gastro-intestinal Symptoms and Mood States in Unprofessional Bodybuilders: A Randomized, Double Blind, Placebo-Controlled Clinical Trial Study

The Effect of Probiotic Supplementation on Gastro-intestinal Symptoms and Mood States in Unprofessional Bodybuilders: A Randomized, Double Blind, Placebo-Controlled Clinical Trial Study | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article The Effect of Probiotic Supplementation on Gastro-intestinal Symptoms and Mood States in Unprofessional Bodybuilders: A Randomized, Double Blind, Placebo-Controlled Clinical Trial Study Nastaran Mahmoudi Shirkoohi, Alireza Chegol, Hamed Mohammadi, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7147381/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 15 You are reading this latest preprint version Abstract Background Emerging evidence suggests that probiotics may alleviate Gastrointestinal (GI) disturbances and positively influence mood by modulating the gut-brain axis. However, there is limited research specifically examining the effects of probiotic supplementation on GI symptoms and mood in bodybuilders. This study aims to investigate the effect of probiotics on Gastrointestinal health and psychological well-being in this population. Method This study was conducted as a randomized, double-blind, placebo-controlled trial involving 36 unprofessional bodybuilders. Participants were randomly assigned to two groups to receive either 2 billion colony-forming units (CFU) of probiotic (n = 18) or a placebo (n = 18) for 12 weeks. The change in symptoms wase determined using the Gastrointestinal Symptom Rating Scale (GSRS) and the Brunel Mood Scale (BRUMS). Results After 12 weeks of probiotic supplementation, athletes experienced a significant reduction in overall GSRS )p = 0.005), reflux (p = 0.04), and abdominal pain (p = 0.007) scores and significantly improved vigor (p = 0.002), peace (p = 0.004), and happiness score (p < 0.001) compared to their baseline measurements. Between-group analysis also revealed a significant reduction in reflux (p = 0.01), abdominal pain (P = 0.01), and overall GSRS scores (P = 0.02). We observed a significant rise in vigor (p = 0.02), peace (p = 0.01), and happiness score (p < 0.001). Conclusion The results demonstrated that probiotic supplementation has the potential to improve Gastrointestinal symptoms and mood states in athletes. Further studies with adequate power should be performed. Trial registration: Iranian Registry of Clinical Trials IRCT20240424061562N1, Registration date: 2024–07–30, Trial Id: 77722. probiotic gastrointestinal symptoms mood states bodybuilders Figures Figure 1 Introductions For bodybuilders, who often experience intense training regimens and specific dietary restrictions, maintaining optimal Gastrointestinal (GI) health is essential for enhancing performance and recovery [ 1 ]. GI disturbances, such as bloating, gas, and diarrhea, are common among athletes and can adversely affect both physical performance and mood [ 2 ]. These issues have been linked to changes in intestinal permeability and weakened intestinal barrier function [ 3 ]. Elevated gastrointestinal permeability, often referred to as "leaky gut syndrome," can result in endotoxemia, which increases vulnerability to infections and autoimmune conditions due to the entry of harmful microorganisms and toxins into the bloodstream [ 4 ]. Approaches aimed at reducing GI injury and inflammation during exercise could lessen abdominal discomfort and enhance the absorption of fluids, electrolytes, and nutrients, ultimately benefiting athletic health, performance, and recovery [ 5 , 6 ]. Moreover, the intense physical stressors associated with bodybuilding can also impact mental well-being, leading to anxiety and mood fluctuations that may affect motivation and performance [ 7 ]. Research suggests that the gut-brain axis _the intricate communication network linking the gut and the brain_ plays a significant role in this dynamic [ 8 ]. This bi-directional communication system between the gut and the brain is influenced by gut microbiota, which can modulate neurotransmitter production, immune responses, and neuroactive substances [ 9 ]. Many recent studies have focused on probiotic supplementation, whether using single or multi-strain products, as a possible solution for enhancing health and athletic performance in athletes [ 6 , 10 , 11 ]. Probiotics may help restore the balance of gut microbiota disrupted by high-intensity training and dietary changes, thereby reducing inflammation and enhancing nutrient absorption [ 12 ]. Also, probiotics may offer a viable solution by promoting a healthy gut microbiome, potentially alleviating GI discomfort while also influencing mood-regulating neurotransmitters [ 13 ]. Despite the promising potential of probiotics, there is limited research specifically addressing their effects on GI symptoms and mood states in bodybuilders. Understanding how probiotic supplementation can impact these factors is critical for optimizing athletic performance and improving quality of life in this population. Therefore, this study aims to investigate the effect of probiotic supplementation on GI symptoms and mood states in bodybuilders, providing valuable insights into the role of gut microbiota management in enhancing athletic performance. MATERIALS AND METHODS Study design The present study was a randomized, placebo-controlled, double‐ blind parallel clinical trial that was conducted on unprofessional bodybuilders from September 2024 to January 2024, which was registered in the Iranian Clinical Trials Registration of clinical trials (IRCT20240424061562N1, Registration date: 2024–07–30, Trial Id: 77722). The study protocol was approved by the Ethics Committee of Tehran University of Medical Sciences (approval code: IR.TUMS.MEDICINE.REC.1403.157) and was conducted according to the Declaration of Helsinki. Also, this study adheres to CONSORT guidelines. This study is designed to be conducted at Sports Medicine Research, affiliated with Tehran University of Medical Sciences, Tehran, Iran. Researchers obtained written informed consent from study subjects before participation in the clinical trial (appendix. 1) . The eligibility criteria included a minimum of 3 months of bodybuilding training experience; healthy adults between 20 and 40 years old who train an average of 6–10 hours per week; and athletes who voluntarily agreed to participate by signing an informed consent form. Exclusion criteria comprised professional athletes training for over 6 hours weekly; pregnant or lactating women; women experiencing premature menopause; and individuals with medical conditions that could hinder their ability to perform or engage in exercise, Participants who frequently consume probiotic products, such as dietary fibers like inulin and galacto-oligosaccharides, on a regular basis for at least one month before and during the study; current smokers; individuals with a suspected or confirmed history of alcohol or drug misuse; those who have consumed food or sports supplements within four to six weeks prior to and during the intervention; individuals following special diets; those who have taken antibiotics during the study or within the past four weeks; individuals with gastrointestinal diseases such as food allergies, celiac disease, or irritable bowel syndrome within the previous two months; participants engaged in sports other than bodybuilding; and those experiencing acute gastrointestinal symptoms within two months prior to the study were excluded from participation. Exclusion criteria included: allergy to studied agents; noncompliance in consumption of the supplements (< 90%); taking antibiotics during the study; and pregnancy during the study. Sample size The sample size was determined to be n = 18 for each group, based on Huang et al. (2018) [ 14 ], taking into account a type one error rate of 5%, a statistical power of 80%, and a dropout rate of 20%. Randomization and blinding Participants were randomly assigned using the permutated block randomization method, with gender-based blocks of 4:6:6. The random allocation was performed via the website: https://www.sealedenvelope.com/simple-randomiser/v1/lists [Accessed 2 Aug 2024]. A trained nutritionist was responsible for enrolling and assigning subjects to the groups. The manufacturer coded the probiotic supplement and its placebo as codes 1 and 2, packing them in identical boxes with the same color, shape, and size. Researchers, outcome assessors, and athletes were kept unaware of the random allocation sequence until data analysis was completed. Study interventions Eligible athletes entered a two-week run-in period after enrollment to assess their dietary intake and physical activity. Additionally, data related to body composition, gastrointestinal symptoms, and mood states were collected. Subsequently, 40 eligible participants were randomly assigned to two groups: the probiotic supplement group (n = 20) and the control group (n = 20). The intervention group daily received a probiotic supplement (2 billion CFU of Lactobacillus acidophilus, Lactobacillus plantarum, and Lactobacillus paracasei ) for 12 weeks, while the control group received a placebo (100 mg maltodextrin). Each supplement was provided to the participants at the beginning of each month and they were advised to store it in refrigeration to preserve bacteria. Additionally, at the end of each month, they were asked to return the empty bottles to the researcher. If any of the participants had not consumed 10% of their capsules, this state was considered as a research dropout and were excluded from the study. Probiotic supplements and the placebo were provided by Tak Gen Zist Pharmaceutical Company, Tehran, Iran. Nutritional assessments To assess the dietary intake of athletes, an appropriate method—specifically a 3-day food record covering two weekdays and one weekend day—will be employed. Participants will complete this record through face-to-face interviews at the beginning, midpoint, and end of the intervention. The nutritional data will be analyzed using Nutritionist IV software (First Databank, San Bruno, CA, USA), which has been adapted for Iranian foods. Participants will also receive a list of foods containing probiotics (such as yogurt, kefir, buttermilk, cheese, milk, kimchi, kombucha tea, tempeh, apple cider vinegar, balsamic vinegar, soy sauce, sourdough bread, dark chocolate, olives, Parmesan cheese, pickles, garlic, onions, feta cheese, smoothies, sauerkraut, mixed pickles, pickled onions, pickled lips, pickled cucumbers, probiotic-containing tablets and mouth fresheners). They will be instructed not to include these products in their diet during the intervention and to maintain their usual diet and physical activity levels. Any changes in dietary intake during the study will be considered in the statistical analysis to control for potential confounding factors. Physical activity assessments This project utilized the short form of the International Physical Activity Questionnaire (IPAQ) to assess physical activity. The questionnaire includes seven brief questions about the duration and number of days per week engaging in heavy or moderate activities, walking, and sitting time. It was used to gather data on non-daily activities over the past week, completed both before and after the intervention. The resulting data are expressed as average MET-minutes per week. In addition to using the IPAQ, a structured exercise program was developed in collaboration with the athletes’ coaches to standardize and control the type and volume of activity among participants. This program comprised specific exercises with defined frequency, type, and intensity to ensure consistency and minimize variability caused by differing routines. Regular oversight of the program’s implementation was maintained through weekly meetings and review of daily exercise logs to confirm participants followed the prescribed regimen. Coaches maintained direct contact with the athletes, monitored their performance, communicated feedback to the researchers (N.M, A.CH), and made necessary adjustments to the training plan as needed. GI symptoms questionnaire The Gastrointestinal Symptom Rating Scale (GSRS), validated in Iran, was used to assess participants' gastrointestinal symptoms at the start and end of the study. It consists of 15 questions scored on a 7-point Likert scale, with higher scores indicating more severe symptoms. The scale evaluates five subdomains: indigestion, constipation, diarrhea, heartburn, and reflux. Each subdomain score is calculated as an average of relevant items, and the overall GSRS score is the average of these five subdomain scores [ 15 ]. Mood states questionnaire The Brums Mood Questionnaire, adapted for Iran, is a 26-item tool used to assess athletes' moods across eight emotional states, divided into positive (vitality, peace, happiness) and negative (tension, depression, anger, fatigue, confusion) subscales. Each item is scored from 0 to 5, with higher scores indicating greater mood disturbance. The scores for each subscale are averaged, and the total score ranges from 0 to 128, reflecting overall mood intensity [ 16 , 17 ]. Statistical analysis Data analysis was conducted using the Statistical Package for the Social Sciences (SPSS) software (version 28; SPSS Inc., Chicago, IL), following an intention-to-treat (ITT) approach, with a per-protocol (PP) analysis performed as necessary. Chi-square tests were utilized to compare qualitative variables between the probiotic and control groups, while independent Student's t-tests were used for quantitative variables between the two groups. The Shapiro–Wilk test assessed the normality of quantitative variable distributions. For within-group and between-group comparisons, paired t-tests and Student's t-tests were applied based on normality, while Wilcoxon and Mann–Whitney U tests were used for non-normally distributed data. Analysis of covariance (ANCOVA) assessed changes between groups for normally distributed variables, adjusting for baseline mean values in each group. Bonferroni correction was applied for multiple testing adjustments. Quade’s ANCOVA was used when the outcome distribution was not normal or when assumptions were violated. Data were presented as means and standard deviations, with a p-value of < 0.05 considered statistically significant. Results While 80 participants were initially recruited for this study, 2 participants from the placebo group and 2 from the probiotic group were excluded due to refusal to participate, antibiotic use, and gastrointestinal complications. Ultimately, 18 athletes completed the trial in each group (Fig. 1). Table 1 indicated baseline characteristics of study patients. The mean ages were 29.65 ± 7.68 years for the probiotic group and 28.10 ± 6.96 years for the placebo group. The chi-square test revealed that both groups comprised 11 males (55%) and 9 females (45%), with no significant differences between them. Table 2 presents the baseline characteristics of the athletes, showing no remarkable differences between the intervention and placebo groups. Additionally, there were no significant differences in total energy and macronutrient intake within or between the two groups. Table 2 Dietary intake of study participants at the baseline and after 6 and 12 weeks of intervention* Variables Time Probiotic (n = 18) Placebo (n = 18) P-value (time*group) Energy (kcal/day) At baseline 1399.9 ± 360.83 1442.88 ± 529.79 At 6 weeks 1473.55 ± 452.57 1493.98 ± 481.17 0.631 At 12 weeks 1486.60 ± 274.57 1478.00 ± 368.77 Carbohydrates (g/day) At baseline 156.94 ± 50.34 155.86 ± 74.26 At 6 weeks 167.58 ± 57.96 170.98 ± 75.79 0.531 At 12 weeks 169.86 ± 50.40 176.65 ± 66.37 Protein (g/day) At baseline 90.32 ± 29.24 95.50 ± 35.18 At 6 weeks 84.14 ± 19.98 98.92 ± 28.13 0.503 At 12 weeks 89.96 ± 22.66 101.58 ± 27.35 Fat (g/day) At baseline 46.24 ± 23.57 48.41 ± 24.79 At 6 weeks 54.12 ± 26.39 45.45 ± 18.74 0.607 At 12 weeks 48.06 ± 17.34 55.52 ± 25.09 *All data are reported as mean ± standard deviation. obtained from repeated measures ANOVA. Table 3 shows changes in the physical activity of bodybuilders before and after supplement intake, which have not resulted in any significant differences (p > 0.05). Table 3 Comparison of physical activity between the two groups receiving probiotic supplements and placebo* variable Time Probiotic supplement Placebo Physical activity (MET/MIN/WEEK) Before 2899.90 ± 582.67 2780.15 ± 584.96 After 3018.85 ± 624.17 2837.15 ± 612.34 Mean changes 118.95 ± 303.85 57 ± 395.16 P-value 0.09 0.52 *All data are reported as mean ± standard deviation. Comparison of within-group changes before and after the study using Paired T-Test. After the trial, the total GSRS score significantly decreased in the probiotic group (p = 0.02), with notable inter-group differences (p = 0.005). There was a near-significant reduction in the indigestion score for the probiotic group compared to the placebo group (p = 0.056). The reflux score in the probiotic group also showed a significant reduction (p = 0.04) and was significantly lower than that of the placebo group (p = 0.01). Additionally, a significant decrease in abdominal pain was observed after probiotic consumption (p = 0.007), with a statistically significant difference between the groups (p = 0.01). However, no significant differences were noted for other gastrointestinal symptoms (Table 4) after adjusting for baseline values. Table 4 The effect of probiotic consumption on gastrointestinal disorders* Variables Time Probiotic (n = 20) Placebo (n = 20) P-value b P-value c Reflux score Before 1.47 ± 0.85 1.40 ± 1.17 0.14 After 1.12 ± 0.39 1.55 ± 1.15 0.07 0.01 Mean changes -0.35 ± 0.72 P-value a 0.04 0.09 Abdominal pain score Before 1.63 ± 1.03 1.36 ± 0.86 0.29 After 1.31 ± 0.63 1.40 ± 0.85 0.73 0.01 Mean changes -0.31 ± 0.43 P-value a 0.007 0.31 Indigestion score Before 1.35 ± 0.61 1.18 ± 0.44 0.50 After 1.22 ± 0.24 1.38 ± 0.38 0.16 0.05 Mean changes -0.12 ± 0.55 P-value a 0.36 0.01 Diarrhea score Before 1.36 ± 0.67 2.05 ± 1.24 0.04 After 1.20 ± 0.34 1.78 ± 1.01 0.04 0.66 Mean changes -0.16 ± 0.42 P-value a 0.07 0.01 Constipation score Before 1.33 ± 0.91 1.23 ± 0.67 0.82 After 1.46 ± 1.05 1.58 ± 1.07 0.34 0.34 Mean changes 0.13 ± 1.06 0.35 ± 0.93 P-value a 0.91 0.08 Total Before 21.35 ± 9.83 21.50 ± 9.34 0.69 After 17.95 ± 4.72 21.15 ± 9.24 0.21 0.02 Mean changes 1.47 ± 0.85 P-value a 1.12 ± 0.39 0.24 *Note. All data are reported as mean ± standard deviation. a. obtained from Wilcoxon. b. obtained from the Mann-Whitney. C. obtained from the Quade’s ANCOVA adjusted based on the baseline value. The effects of probiotic consumption on mood states are summarized in Table 5. Within-group comparisons revealed significant improvements in vigor (p = 0.02), peace (p = 0.004), and happiness (p < 0.001). When compared to the placebo group, probiotic consumption also resulted in significant increases in vigor (p = 0.02), peace (p = 0.01), and happiness (p < 0.001). However, there were no statistically significant differences observed for other mood symptoms. Table 5 The effect of probiotic consumption on mood states. Variables Time Probiotic (n = 20) Placebo (n = 20) P-value b P-value c Tension score Before 5.20 ± 3.99 5.25 ± 3.56 0.96 After 5.22 ± 2.96 5.52 ± 3.23 0.75 0.50 Mean changes 0.02 ± 1.99 0.27 ± 0.97 P-value a 0.96 0.22 Depression score Before 3.40 ± 3.15 5.00 ± 3.00 0.10 After 3.44 ± 3.04 4.70 ± 2.27 0.14 0.58 Mean changes 0.04 ± 3.01 -0.30 ± 1.89 P-value a 0.95 0.48 Anger score Before 4.15 ± 3.26 3.15 ± 2.23 0.26 After 3.86 ± 2.45 3.41 ± 1.72 0.50 0.63 Mean changes -0.28 ± 1.78 0.26 ± 1.30 P-value a 0.48 0.37 Vigor score Before 10.30 ± 2.17 8.25 ± 2.07 0.004 After 10.88 ± 2.19 8.22 ± 2.06 < 0.001 0.02 Mean changes 0.58 ± 1.01 -0.02 ± 0.99 P-value a 0.02 0.91 Fatigue score Before 4.95 ± 3.72 5.15 ± 2.39 0.84 After 4.64 ± 3.35 5.01 ± 2.05 0.67 0.64 Mean changes -0.30 ± 1.91 -0.13 ± 1.27 P-value a 0.48 0.64 Confusion score Before 5.15 ± 3.92 5.60 ± 3.06 0.68 After 5.06 ± 2.87 5.81 ± 2.72 0.40 0.41 Mean changes -0.08 ± 2.88 0.21 ± 1.23 P-value a 0.89 0.45 Calmness score Before 6.80 ± 2.98 7.20 ± 2.72 0.66 After 8.08 ± 2.47 7.02 ± 2.17 0.15 0.01 Mean changes 1.28 ± 1.77 -0.18 ± 2.15 P-value a 0.004 0.71 Happiness score Before 8.60 ± 2.37 7.50 ± 2.94 0.20 After 10.01 ± 2.30 7.45 ± 2.41 < 0.001 < 0.001 Mean changes 1.41 ± 1.33 -0.05 ± 1.45 P-value a < 0.001 0.88 Total Before 48.55 ± 13.07 47.10 ± 7.24 0.66 After 51.20 ± 9.32 47.15 ± 7.02 0.12 0.06 Mean changes 2.65 ± 8.48 0.05 ± 4.05 P-value a 0.17 0.95 *Note. All data are reported as mean ± standard deviation. a. obtained from paired t-test. b. obtained from the Independent T Test. C. obtained from ANCOVA adjusted based on the baseline value. It is worth mentioning that during the intervention period, no symptoms or risks occurred for the participants, except for one individual in the intervention group who experienced severe diarrhea and refused to continue with the study. Discussion Research on the effects of probiotic supplementation in athletes and its implications in clinical settings is limited. To our knowledge, this investigation is the first double-blind, placebo-controlled randomized clinical trial (RCT) to explore the effects of probiotic supplementation on GI symptoms and mood states specifically in unprofessional bodybuilders. The key findings from this trial indicate that, after three months of probiotic supplementation, there were significant reductions in reflux, abdominal pain, and overall GSRS scores among the athletes. Additionally, the probiotic group experienced increases in scores for vigor, calmness, and happiness compared to the placebo group. Supporting the present finding, in a recent RCT performed by Wang et al, showed that, probiotics have the potential to improve athletic performance by optimizing the balance of gut microbiota, and alleviating GI symptoms [ 18 ]. In the study by Roberts et al., the average symptom severity was significantly lower in the probiotic group after months 1 and 3 of intervention than in the placebo group [ 19 ]. Additionally, West et al. found that the self-reported severity score for gastrointestinal illnesses in males taking probiotics was, on average, 0.7 points lower on the scale compared to males receiving the placebo. Furthermore, this beneficial effect of the probiotic became more pronounced as the training load increased [ 20 ]. In opposition to our findings, Pugh et al. observed no notable differences in the total symptom severity score of GI during the composition in 14 days of supplementation with probiotics among trained cyclists [ 21 ]. It should be mentioned that the type of exercise, duration of intervention, and the intensity of the workout, which affect GI symptoms, differ from our study. In terms of mood states, Noorifard et al. reported that 12 weeks of supplementation with probiotic juice has beneficial effects on the body's antioxidant system and mental health parameters in athlete soldiers [ 22 ]. In contrast, Salleh et al. found that probiotic supplementation had no impact on the mood of badminton players [ 23 ]. Exercise can potentially enhance gut permeability, enabling toxins to pass from the gut into the bloodstream [ 24 ]. This increased permeability is often associated with higher levels of bacterial endotoxins, which coincide with elevated inflammatory cytokines [ 25 ]. Therefore, it has been proposed that strengthening gut barrier function through probiotic supplementation could help reduce exercise-induced endotoxemia and inflammation, as well as potentially prevent gastrointestinal issues [ 26 – 28 ]. In another hand, the gut-associated lymphoid tissue (GALT) plays a crucial role in immune function [ 29 ]. Probiotics can enhance the production of anti-inflammatory cytokines while reducing pro-inflammatory cytokines, which helps to maintain an appropriate immune balance[ 30 ]. By promoting a balanced immune response, probiotics can help reduce the incidence of GI infections and inflammation, which are common issues faced by athletes [ 31 ]. This immune modulation may also contribute to a reduction in exercise-induced GI symptoms, allowing bodybuilders to train more effectively without the burden of digestive discomfort [ 32 ]. Moreover, the reduction in inflammation through probiotic use may also contribute to improved mental health outcomes, as chronic inflammation is linked to mood disorders [ 33 ]. The gut-brain axis is a critical pathway through which probiotics exert their effects on mental health. Probiotics can influence the production of neurotransmitters and modulate the immune response, which may help reduce symptoms of depression and mental stress in athletes [ 34 ]. The gut microbiota may help lower corticosterone release triggered by stress, thereby alleviating anxiety and depression-related behaviors by enhancing the expression of gamma-aminobutyric acid (GABA) receptors in the brain [ 35 ]. Probiotics might also influence gut functions by activating gut-brain axis responses, potentially converting dopamine into noradrenaline within both the central and peripheral nervous systems. This transformation could result in a reduction of depressive behaviors [ 36 ]. In addition, Probiotics contribute to the reduction of oxidative stress, which is linked to improved mental health outcomes. This is evidenced by increased antioxidant levels in athletes consuming probiotics, which correlates with better psychological parameters [ 37 ]. This study has several notable strengths, most importantly being conducted for the first time in this context. It also employed stratified permuted block randomization and a double-blind, placebo-controlled design, which enhanced the reliability of the results. Additionally, the findings could provide practical guidance for bodybuilders aiming to boost their gut health and overall performance. Nonetheless, there are some limitations to consider when interpreting these results. The study may not fully account for individual differences in response to probiotics, such as genetic factors or variations in preexisting gut microbiota, which can greatly affect outcomes. Furthermore, the reliance on self-reported data for adherence and physical activity could introduce recall bias and reduce the accuracy of the findings. Future research with larger, well-controlled studies is necessary to establish more conclusive evidence. Conclusion In summary, probiotics offer a multifaceted approach to managing GI symptoms and mood states in bodybuilders through various mechanisms. Given the unique dietary and training challenges faced by bodybuilders, incorporating probiotics into their regimen may provide significant benefits for GI health and mental states, ultimately supporting their performance and recovery. As with any supplement, careful consideration and individualized approaches are essential for achieving the best outcomes. Abbreviations GI; Gastrointestinal, GSRS; Gastrointestinal Symptom Rating Scale, BRUMS; Brunel Mood Scale, IPAQ; International Physical Activity Questionnaire, GALT; gut-associated lymphoid tissue, GABA; gamma-aminobutyric acid, RCT; randomized clinical trial Declarations Ethics approval and consent to participate Written informed consent was obtained from participants before participation in the research project. The current trial received approval from Medical Ethics Committee of Tehran University of Medical Sciences, Tehran, Iran (IR. TUMS. MEDICNE.REC.1403.157). This study was conducted in accordance with the Declaration of Helsinki. Consent for publication Not applicable. Data availability All data generated or analyzed during this study are included in this published article. Competing interests The authors declare no competing interests. Funding The trial funding was supported by Tehran University of Medical Science (Grant number: 72295). The funder is not involved in the study design, collection, management, analysis and interpretation, writing of the manuscript, and the decision to submit the report for publication, including whether they will have ultimate authority over any of these activities. Author’s contributions Nastaran Mahmoudi Shirkoohi: Methodology; writing – original draft; writing – review and editing. Alireza Chegol: Writing – original draft; writing – review and editing. Hamed Mohammadi: Methodology; software; writing – original draft; writing – review and editing. Sakineh Shab-bidar: Methodology; validation; Formal analysis; visualization; writing – review and editing. Siroos Choobineh: Methodology; writing – review and editing. Kurosh Djafarian: Conceptualization; methodology; project administration; resources; writing – review and editing. Acknowledgments We sincerely thank the Tak Gen Zist Pharmaceutical Company for their support. References Maughan, R.J. and S.M. 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Noorifard, M., et al., Effect of probiotic supplementation on oxidative stress enzymes and mental health of athletes. Annals of Military and Health Sciences Research, 2019. 17 (1). Salleh, R.M., et al., Effects of probiotics on anxiety, stress, mood and fitness of badminton players. Nutrients, 2021. 13 (6): p. 1783. Criswell, D.S., et al., Chronic exercise and the pro-inflammatory response to endotoxin in the serum and heart. Immunology letters, 2004. 95 (2): p. 213-220. Fukui, H., Increased intestinal permeability and decreased barrier function: does it really influence the risk of inflammation? Inflammatory intestinal diseases, 2016. 1 (3): p. 135-145. Cinca-Morros, S. and J. Álvarez-Herms, The Importance of Maintaining and Improving a Healthy Gut Microbiota in Athletes as a Preventive Strategy to Improve Heat Tolerance and Acclimatization. Microorganisms, 2024. 12 (6): p. 1160. Lennon, S., The effects of probiotic supplementation on endurance athletes: Influences on gut dysfunction, microbiome, and gut discomfort . 2022: The University of Memphis. Smith, K.A., et al., Gastrointestinal pathophysiology during endurance exercise: endocrine, microbiome, and nutritional influences. European Journal of Applied Physiology, 2021. 121 (10): p. 2657-2674. Mörbe, U.M., et al., Human gut-associated lymphoid tissues (GALT); diversity, structure, and function. Mucosal immunology, 2021. 14 (4): p. 793-802. Starkie, R., et al., Circulating monocytes are not the source of elevations in plasma IL-6 and TNF-α levels after prolonged running. American Journal of Physiology-Cell Physiology, 2001. 280 (4): p. C769-C774. Łagowska, K. and J. Bajerska, Probiotic supplementation and respiratory infection and immune function in athletes: systematic review and meta-analysis of randomized controlled trials. Journal of Athletic Training, 2021. 56 (11): p. 1213-1223. Wosinska, L., et al., The potential impact of probiotics on the gut microbiome of athletes. Nutrients, 2019. 11 (10): p. 2270. Di Dio, M., et al., Effects of Probiotic supplementation on sports performance and performance-related features in athletes: a systematic review. International Journal of Environmental Research and Public Health, 2023. 20 (3): p. 2226. Sivamaruthi, B.S., P. Kesika, and C. Chaiyasut, Effect of probiotics supplementations on health status of athletes. International Journal of Environmental Research and Public Health, 2019. 16 (22): p. 4469. Bravo, J.A., et al., Ingestion of Lactobacillus strain regulates emotional behavior and central GABA receptor expression in a mouse via the vagus nerve. Proceedings of the National Academy of Sciences, 2011. 108 (38): p. 16050-16055. Pinto-Sanchez, M.I., et al., Probiotic Bifidobacterium longum NCC3001 reduces depression scores and alters brain activity: a pilot study in patients with irritable bowel syndrome. Gastroenterology, 2017. 153 (2): p. 448-459. e8. Zhang, L., R. Zhang, and L. Li, Effects of probiotic supplementation on exercise and the underlying mechanisms. Foods, 2023. 12 (9): p. 1787. Table 1 Table 1 is available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files table1.docx appendix1.docx CONSORT2010Checklist.doc Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 06 Feb, 2026 Reviews received at journal 06 Feb, 2026 Reviews received at journal 04 Feb, 2026 Reviews received at journal 08 Jan, 2026 Reviews received at journal 26 Dec, 2025 Reviewers agreed at journal 20 Dec, 2025 Reviewers agreed at journal 18 Dec, 2025 Reviewers agreed at journal 18 Dec, 2025 Reviews received at journal 05 Oct, 2025 Reviewers agreed at journal 01 Oct, 2025 Reviewers invited by journal 16 Sep, 2025 Editor assigned by journal 05 Aug, 2025 Editor invited by journal 04 Aug, 2025 Submission checks completed at journal 03 Aug, 2025 First submitted to journal 31 Jul, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7147381","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":518379606,"identity":"7f0d6a0f-8bee-407e-ab16-f4dff3205e2c","order_by":0,"name":"Nastaran Mahmoudi Shirkoohi","email":"","orcid":"","institution":"Tehran University of medical science","correspondingAuthor":false,"prefix":"","firstName":"Nastaran","middleName":"Mahmoudi","lastName":"Shirkoohi","suffix":""},{"id":518379607,"identity":"8f23c6ed-acb3-4473-b1a9-2079dd147990","order_by":1,"name":"Alireza Chegol","email":"","orcid":"","institution":"Tehran University of 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GI disturbances, such as bloating, gas, and diarrhea, are common among athletes and can adversely affect both physical performance and mood [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. These issues have been linked to changes in intestinal permeability and weakened intestinal barrier function [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Elevated gastrointestinal permeability, often referred to as \"leaky gut syndrome,\" can result in endotoxemia, which increases vulnerability to infections and autoimmune conditions due to the entry of harmful microorganisms and toxins into the bloodstream [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Approaches aimed at reducing GI injury and inflammation during exercise could lessen abdominal discomfort and enhance the absorption of fluids, electrolytes, and nutrients, ultimately benefiting athletic health, performance, and recovery [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eMoreover, the intense physical stressors associated with bodybuilding can also impact mental well-being, leading to anxiety and mood fluctuations that may affect motivation and performance [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Research suggests that the gut-brain axis _the intricate communication network linking the gut and the brain_ plays a significant role in this dynamic [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. This bi-directional communication system between the gut and the brain is influenced by gut microbiota, which can modulate neurotransmitter production, immune responses, and neuroactive substances [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eMany recent studies have focused on probiotic supplementation, whether using single or multi-strain products, as a possible solution for enhancing health and athletic performance in athletes [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Probiotics may help restore the balance of gut microbiota disrupted by high-intensity training and dietary changes, thereby reducing inflammation and enhancing nutrient absorption [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Also, probiotics may offer a viable solution by promoting a healthy gut microbiome, potentially alleviating GI discomfort while also influencing mood-regulating neurotransmitters [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eDespite the promising potential of probiotics, there is limited research specifically addressing their effects on GI symptoms and mood states in bodybuilders. Understanding how probiotic supplementation can impact these factors is critical for optimizing athletic performance and improving quality of life in this population. Therefore, this study aims to investigate the effect of probiotic supplementation on GI symptoms and mood states in bodybuilders, providing valuable insights into the role of gut microbiota management in enhancing athletic performance.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003e\u003cb\u003eStudy design\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe present study was a randomized, placebo-controlled, double‐ blind parallel clinical trial that was conducted on unprofessional bodybuilders from September 2024 to January 2024, which was registered in the Iranian Clinical Trials Registration of clinical trials (IRCT20240424061562N1, Registration date: 2024\u0026ndash;07\u0026ndash;30, Trial Id: 77722). The study protocol was approved by the Ethics Committee of Tehran University of Medical Sciences (approval code: IR.TUMS.MEDICINE.REC.1403.157) and was conducted according to the Declaration of Helsinki. Also, this study adheres to CONSORT guidelines. This study is designed to be conducted at Sports Medicine Research, affiliated with Tehran University of Medical Sciences, Tehran, Iran. Researchers obtained written informed consent from study subjects before participation in the clinical trial \u003cb\u003e(appendix. 1)\u003c/b\u003e. The eligibility criteria included a minimum of 3 months of bodybuilding training experience; healthy adults between 20 and 40 years old who train an average of 6\u0026ndash;10 hours per week; and athletes who voluntarily agreed to participate by signing an informed consent form. Exclusion criteria comprised professional athletes training for over 6 hours weekly; pregnant or lactating women; women experiencing premature menopause; and individuals with medical conditions that could hinder their ability to perform or engage in exercise, Participants who frequently consume probiotic products, such as dietary fibers like inulin and galacto-oligosaccharides, on a regular basis for at least one month before and during the study; current smokers; individuals with a suspected or confirmed history of alcohol or drug misuse; those who have consumed food or sports supplements within four to six weeks prior to and during the intervention; individuals following special diets; those who have taken antibiotics during the study or within the past four weeks; individuals with gastrointestinal diseases such as food allergies, celiac disease, or irritable bowel syndrome within the previous two months; participants engaged in sports other than bodybuilding; and those experiencing acute gastrointestinal symptoms within two months prior to the study were excluded from participation.\u003c/p\u003e\u003cp\u003eExclusion criteria included: allergy to studied agents; noncompliance in consumption of the supplements (\u0026lt;\u0026thinsp;90%); taking antibiotics during the study; and pregnancy during the study.\u003c/p\u003e\u003cp\u003e\u003cb\u003eSample size\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe sample size was determined to be n\u0026thinsp;=\u0026thinsp;18 for each group, based on Huang et al. (2018) [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], taking into account a type one error rate of 5%, a statistical power of 80%, and a dropout rate of 20%.\u003c/p\u003e\u003cp\u003e\u003cb\u003eRandomization and blinding\u003c/b\u003e\u003c/p\u003e\u003cp\u003eParticipants were randomly assigned using the permutated block randomization method, with gender-based blocks of 4:6:6. The random allocation was performed via the website: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.sealedenvelope.com/simple-randomiser/v1/lists\u003c/span\u003e\u003cspan address=\"https://www.sealedenvelope.com/simple-randomiser/v1/lists\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e [Accessed 2 Aug 2024]. A trained nutritionist was responsible for enrolling and assigning subjects to the groups. The manufacturer coded the probiotic supplement and its placebo as codes 1 and 2, packing them in identical boxes with the same color, shape, and size. Researchers, outcome assessors, and athletes were kept unaware of the random allocation sequence until data analysis was completed.\u003c/p\u003e\u003cp\u003e\u003cb\u003eStudy interventions\u003c/b\u003e\u003c/p\u003e\u003cp\u003eEligible athletes entered a two-week run-in period after enrollment to assess their dietary intake and physical activity. Additionally, data related to body composition, gastrointestinal symptoms, and mood states were collected. Subsequently, 40 eligible participants were randomly assigned to two groups: the probiotic supplement group (n\u0026thinsp;=\u0026thinsp;20) and the control group (n\u0026thinsp;=\u0026thinsp;20). The intervention group daily received a probiotic supplement (2\u0026nbsp;billion CFU of \u003cem\u003eLactobacillus acidophilus, Lactobacillus plantarum, and Lactobacillus paracasei\u003c/em\u003e) for 12 weeks, while the control group received a placebo (100 mg maltodextrin). Each supplement was provided to the participants at the beginning of each month and they were advised to store it in refrigeration to preserve bacteria. Additionally, at the end of each month, they were asked to return the empty bottles to the researcher. If any of the participants had not consumed 10% of their capsules, this state was considered as a research dropout and were excluded from the study. Probiotic supplements and the placebo were provided by Tak Gen Zist Pharmaceutical Company, Tehran, Iran.\u003c/p\u003e\u003cp\u003e\u003cb\u003eNutritional assessments\u003c/b\u003e\u003c/p\u003e\u003cp\u003eTo assess the dietary intake of athletes, an appropriate method\u0026mdash;specifically a 3-day food record covering two weekdays and one weekend day\u0026mdash;will be employed. Participants will complete this record through face-to-face interviews at the beginning, midpoint, and end of the intervention. The nutritional data will be analyzed using Nutritionist IV software (First Databank, San Bruno, CA, USA), which has been adapted for Iranian foods. Participants will also receive a list of foods containing probiotics (such as yogurt, kefir, buttermilk, cheese, milk, kimchi, kombucha tea, tempeh, apple cider vinegar, balsamic vinegar, soy sauce, sourdough bread, dark chocolate, olives, Parmesan cheese, pickles, garlic, onions, feta cheese, smoothies, sauerkraut, mixed pickles, pickled onions, pickled lips, pickled cucumbers, probiotic-containing tablets and mouth fresheners). They will be instructed not to include these products in their diet during the intervention and to maintain their usual diet and physical activity levels. Any changes in dietary intake during the study will be considered in the statistical analysis to control for potential confounding factors.\u003c/p\u003e\u003cp\u003e\u003cb\u003ePhysical activity assessments\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThis project utilized the short form of the International Physical Activity Questionnaire (IPAQ) to assess physical activity. The questionnaire includes seven brief questions about the duration and number of days per week engaging in heavy or moderate activities, walking, and sitting time. It was used to gather data on non-daily activities over the past week, completed both before and after the intervention. The resulting data are expressed as average MET-minutes per week. In addition to using the IPAQ, a structured exercise program was developed in collaboration with the athletes\u0026rsquo; coaches to standardize and control the type and volume of activity among participants. This program comprised specific exercises with defined frequency, type, and intensity to ensure consistency and minimize variability caused by differing routines. Regular oversight of the program\u0026rsquo;s implementation was maintained through weekly meetings and review of daily exercise logs to confirm participants followed the prescribed regimen. Coaches maintained direct contact with the athletes, monitored their performance, communicated feedback to the researchers (N.M, A.CH), and made necessary adjustments to the training plan as needed.\u003c/p\u003e\u003cp\u003e\u003cb\u003eGI symptoms questionnaire\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe Gastrointestinal Symptom Rating Scale (GSRS), validated in Iran, was used to assess participants' gastrointestinal symptoms at the start and end of the study. It consists of 15 questions scored on a 7-point Likert scale, with higher scores indicating more severe symptoms. The scale evaluates five subdomains: indigestion, constipation, diarrhea, heartburn, and reflux. Each subdomain score is calculated as an average of relevant items, and the overall GSRS score is the average of these five subdomain scores [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003cb\u003eMood states questionnaire\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe Brums Mood Questionnaire, adapted for Iran, is a 26-item tool used to assess athletes' moods across eight emotional states, divided into positive (vitality, peace, happiness) and negative (tension, depression, anger, fatigue, confusion) subscales. Each item is scored from 0 to 5, with higher scores indicating greater mood disturbance. The scores for each subscale are averaged, and the total score ranges from 0 to 128, reflecting overall mood intensity [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eStatistical analysis\u003c/h2\u003e\u003cp\u003eData analysis was conducted using the Statistical Package for the Social Sciences (SPSS) software (version 28; SPSS Inc., Chicago, IL), following an intention-to-treat (ITT) approach, with a per-protocol (PP) analysis performed as necessary. Chi-square tests were utilized to compare qualitative variables between the probiotic and control groups, while independent Student's t-tests were used for quantitative variables between the two groups. The Shapiro\u0026ndash;Wilk test assessed the normality of quantitative variable distributions. For within-group and between-group comparisons, paired t-tests and Student's t-tests were applied based on normality, while Wilcoxon and Mann\u0026ndash;Whitney U tests were used for non-normally distributed data. Analysis of covariance (ANCOVA) assessed changes between groups for normally distributed variables, adjusting for baseline mean values in each group. Bonferroni correction was applied for multiple testing adjustments. Quade\u0026rsquo;s ANCOVA was used when the outcome distribution was not normal or when assumptions were violated. Data were presented as means and standard deviations, with a p-value of \u0026lt;\u0026thinsp;0.05 considered statistically significant.\u003c/p\u003e\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eWhile 80 participants were initially recruited for this study, 2 participants from the placebo group and 2 from the probiotic group were excluded due to refusal to participate, antibiotic use, and gastrointestinal complications. Ultimately, 18 athletes completed the trial in each group (Fig.\u0026nbsp;1). Table\u0026nbsp;1 indicated baseline characteristics of study patients. The mean ages were 29.65 ± 7.68 years for the probiotic group and 28.10 ± 6.96 years for the placebo group. The chi-square test revealed that both groups comprised 11 males (55%) and 9 females (45%), with no significant differences between them. Table\u0026nbsp;2 presents the baseline characteristics of the athletes, showing no remarkable differences between the intervention and placebo groups. Additionally, there were no significant differences in total energy and macronutrient intake within or between the two groups.\u003c/p\u003e\n\u003cdiv\u003e\n\u003c/div\u003e\n\u003cdiv\u003e\n \u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv\u003eTable 2\u003c/div\u003e\n \u003cdiv\u003e\n \u003cp\u003eDietary intake of study participants at the baseline and after 6 and 12 weeks of intervention*\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eVariables\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTime\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eProbiotic\u003c/p\u003e\n \u003cp\u003e(n = 18)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePlacebo\u003c/p\u003e\n \u003cp\u003e(n = 18)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003cp\u003e(time*group)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003e\u003cstrong\u003eEnergy (kcal/day)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAt baseline\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1399.9 ± 360.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1442.88 ± 529.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAt 6 weeks\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1473.55 ± 452.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1493.98 ± 481.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.631\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAt 12 weeks\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1486.60 ± 274.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1478.00 ± 368.77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003e\u003cstrong\u003eCarbohydrates\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(g/day)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAt baseline\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e156.94 ± 50.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e155.86 ± 74.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAt 6 weeks\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e167.58 ± 57.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e170.98 ± 75.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.531\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAt 12 weeks\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e169.86 ± 50.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e176.65 ± 66.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003e\u003cstrong\u003eProtein\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(g/day)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAt baseline\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e90.32 ± 29.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e95.50 ± 35.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAt 6 weeks\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e84.14 ± 19.98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e98.92 ± 28.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.503\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAt 12 weeks\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e89.96 ± 22.66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e101.58 ± 27.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003e\u003cstrong\u003eFat\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(g/day)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAt baseline\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e46.24 ± 23.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e48.41 ± 24.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAt 6 weeks\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e54.12 ± 26.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e45.45 ± 18.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.607\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAt 12 weeks\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e48.06 ± 17.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e55.52 ± 25.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003e*All data are reported as mean ± standard deviation.\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003eobtained from repeated measures ANOVA.\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eTable\u0026nbsp;3 shows changes in the physical activity of bodybuilders before and after supplement intake, which have not resulted in any significant differences (p \u0026gt; 0.05).\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv\u003eTable 3\u003c/div\u003e\n \u003cdiv\u003e\n \u003cp\u003eComparison of physical activity between the two groups receiving probiotic supplements and placebo*\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003evariable\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTime\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eProbiotic supplement\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePlacebo\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003e\u003cstrong\u003ePhysical activity\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(MET/MIN/WEEK)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eBefore\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2899.90 ± 582.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2780.15 ± 584.96\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eAfter\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3018.85 ± 624.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2837.15 ± 612.34\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eMean changes\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e118.95 ± 303.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e57 ± 395.16\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eP-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.52\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003e*All data are reported as mean ± standard deviation.\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003eComparison of within-group changes before and after the study using Paired T-Test.\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eAfter the trial, the total GSRS score significantly decreased in the probiotic group (p = 0.02), with notable inter-group differences (p = 0.005). There was a near-significant reduction in the indigestion score for the probiotic group compared to the placebo group (p = 0.056). The reflux score in the probiotic group also showed a significant reduction (p = 0.04) and was significantly lower than that of the placebo group (p = 0.01). Additionally, a significant decrease in abdominal pain was observed after probiotic consumption (p = 0.007), with a statistically significant difference between the groups (p = 0.01). However, no significant differences were noted for other gastrointestinal symptoms (Table\u0026nbsp;4) after adjusting for baseline values.\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable id=\"Tab4\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv\u003eTable 4\u003c/div\u003e\n \u003cdiv\u003e\n \u003cp\u003eThe effect of probiotic consumption on gastrointestinal disorders*\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eVariables\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTime\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eProbiotic\u003c/p\u003e\n \u003cp\u003e(n = 20)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePlacebo\u003c/p\u003e\n \u003cp\u003e(n = 20)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eReflux score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.47 ± 0.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.40 ± 1.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.12 ± 0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.55 ± 1.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.01\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-0.35 ± 0.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.04\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAbdominal pain score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.63 ± 1.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.36 ± 0.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.31 ± 0.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.40 ± 0.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.01\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-0.31 ± 0.43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.007\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIndigestion score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.35 ± 0.61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.18 ± 0.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.22 ± 0.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.38 ± 0.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-0.12 ± 0.55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.01\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDiarrhea score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.36 ± 0.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.05 ± 1.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.04\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.20 ± 0.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.78 ± 1.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.04\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.66\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-0.16 ± 0.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.01\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eConstipation score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.33 ± 0.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.23 ± 0.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.46 ± 1.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.58 ± 1.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.34\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.13 ± 1.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.35 ± 0.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTotal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21.35 ± 9.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21.50 ± 9.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.95 ± 4.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21.15 ± 9.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.02\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.47 ± 0.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.12 ± 0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\u003cstrong\u003e*Note. All data are reported as mean ± standard deviation.\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\u003cstrong\u003ea. obtained from Wilcoxon.\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\u003cstrong\u003eb. obtained from the Mann-Whitney.\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\u003cstrong\u003eC. obtained from the Quade’s ANCOVA adjusted based on the baseline value.\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eThe effects of probiotic consumption on mood states are summarized in Table 5. Within-group comparisons revealed significant improvements in vigor (p = 0.02), peace (p = 0.004), and happiness (p \u0026lt; 0.001). When compared to the placebo group, probiotic consumption also resulted in significant increases in vigor (p = 0.02), peace (p = 0.01), and happiness (p \u0026lt; 0.001). However, there were no statistically significant differences observed for other mood symptoms.\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable id=\"Tab5\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv\u003eTable 5\u003c/div\u003e\n \u003cdiv\u003e\n \u003cp\u003eThe effect of probiotic consumption on mood states.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eVariables\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTime\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eProbiotic (n = 20)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePlacebo\u003c/p\u003e\n \u003cp\u003e(n = 20)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTension score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.20 ± 3.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.25 ± 3.56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.22 ± 2.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.52 ± 3.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.50\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.02 ± 1.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.27 ± 0.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDepression score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.40 ± 3.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.00 ± 3.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.44 ± 3.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.70 ± 2.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.04 ± 3.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-0.30 ± 1.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAnger score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.15 ± 3.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.15 ± 2.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.86 ± 2.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.41 ± 1.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.63\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-0.28 ± 1.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.26 ± 1.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eVigor score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.30 ± 2.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.25 ± 2.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.004\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.88 ± 2.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.22 ± 2.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt; 0.001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.02\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.58 ± 1.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-0.02 ± 0.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.02\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFatigue score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.95 ± 3.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.15 ± 2.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.64 ± 3.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.01 ± 2.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.64\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-0.30 ± 1.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-0.13 ± 1.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eConfusion score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.15 ± 3.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.60 ± 3.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.06 ± 2.87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.81 ± 2.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.41\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-0.08 ± 2.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.21 ± 1.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCalmness score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.80 ± 2.98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.20 ± 2.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.08 ± 2.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.02 ± 2.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.01\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.28 ± 1.77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-0.18 ± 2.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.004\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHappiness score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.60 ± 2.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.50 ± 2.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.01 ± 2.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.45 ± 2.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt; 0.001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt; 0.001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.41 ± 1.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-0.05 ± 1.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt; 0.001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTotal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBefore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e48.55 ± 13.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e47.10 ± 7.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e51.20 ± 9.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e47.15 ± 7.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.06\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean changes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.65 ± 8.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.05 ± 4.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP-value\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\u003cstrong\u003e*Note. All data are reported as mean ± standard deviation.\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\u003cstrong\u003ea. obtained from paired t-test.\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\u003cstrong\u003eb. obtained from the Independent T Test.\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\u003cstrong\u003eC. obtained from ANCOVA adjusted based on the baseline value.\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eIt is worth mentioning that during the intervention period, no symptoms or risks occurred for the participants, except for one individual in the intervention group who experienced severe diarrhea and refused to continue with the study.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eResearch on the effects of probiotic supplementation in athletes and its implications in clinical settings is limited. To our knowledge, this investigation is the first double-blind, placebo-controlled randomized clinical trial (RCT) to explore the effects of probiotic supplementation on GI symptoms and mood states specifically in unprofessional bodybuilders. The key findings from this trial indicate that, after three months of probiotic supplementation, there were significant reductions in reflux, abdominal pain, and overall GSRS scores among the athletes. Additionally, the probiotic group experienced increases in scores for vigor, calmness, and happiness compared to the placebo group.\u003c/p\u003e\u003cp\u003eSupporting the present finding, in a recent RCT performed by Wang et al, showed that, probiotics have the potential to improve athletic performance by optimizing the balance of gut microbiota, and alleviating GI symptoms [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. In the study by Roberts et al., the average symptom severity was significantly lower in the probiotic group after months 1 and 3 of intervention than in the placebo group [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Additionally, West et al. found that the self-reported severity score for gastrointestinal illnesses in males taking probiotics was, on average, 0.7 points lower on the scale compared to males receiving the placebo. Furthermore, this beneficial effect of the probiotic became more pronounced as the training load increased [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn opposition to our findings, Pugh et al. observed no notable differences in the total symptom severity score of GI during the composition in 14 days of supplementation with probiotics among trained cyclists [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. It should be mentioned that the type of exercise, duration of intervention, and the intensity of the workout, which affect GI symptoms, differ from our study.\u003c/p\u003e\u003cp\u003eIn terms of mood states, Noorifard et al. reported that 12 weeks of supplementation with probiotic juice has beneficial effects on the body's antioxidant system and mental health parameters in athlete soldiers [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. In contrast, Salleh et al. found that probiotic supplementation had no impact on the mood of badminton players [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eExercise can potentially enhance gut permeability, enabling toxins to pass from the gut into the bloodstream [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. This increased permeability is often associated with higher levels of bacterial endotoxins, which coincide with elevated inflammatory cytokines [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Therefore, it has been proposed that strengthening gut barrier function through probiotic supplementation could help reduce exercise-induced endotoxemia and inflammation, as well as potentially prevent gastrointestinal issues [\u003cspan additionalcitationids=\"CR27\" citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. In another hand, the gut-associated lymphoid tissue (GALT) plays a crucial role in immune function [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Probiotics can enhance the production of anti-inflammatory cytokines while reducing pro-inflammatory cytokines, which helps to maintain an appropriate immune balance[\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. By promoting a balanced immune response, probiotics can help reduce the incidence of GI infections and inflammation, which are common issues faced by athletes [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. This immune modulation may also contribute to a reduction in exercise-induced GI symptoms, allowing bodybuilders to train more effectively without the burden of digestive discomfort [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Moreover, the reduction in inflammation through probiotic use may also contribute to improved mental health outcomes, as chronic inflammation is linked to mood disorders [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. The gut-brain axis is a critical pathway through which probiotics exert their effects on mental health. Probiotics can influence the production of neurotransmitters and modulate the immune response, which may help reduce symptoms of depression and mental stress in athletes [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. The gut microbiota may help lower corticosterone release triggered by stress, thereby alleviating anxiety and depression-related behaviors by enhancing the expression of gamma-aminobutyric acid (GABA) receptors in the brain [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Probiotics might also influence gut functions by activating gut-brain axis responses, potentially converting dopamine into noradrenaline within both the central and peripheral nervous systems. This transformation could result in a reduction of depressive behaviors [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. In addition, Probiotics contribute to the reduction of oxidative stress, which is linked to improved mental health outcomes. This is evidenced by increased antioxidant levels in athletes consuming probiotics, which correlates with better psychological parameters [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThis study has several notable strengths, most importantly being conducted for the first time in this context. It also employed stratified permuted block randomization and a double-blind, placebo-controlled design, which enhanced the reliability of the results. Additionally, the findings could provide practical guidance for bodybuilders aiming to boost their gut health and overall performance. Nonetheless, there are some limitations to consider when interpreting these results. The study may not fully account for individual differences in response to probiotics, such as genetic factors or variations in preexisting gut microbiota, which can greatly affect outcomes. Furthermore, the reliance on self-reported data for adherence and physical activity could introduce recall bias and reduce the accuracy of the findings. Future research with larger, well-controlled studies is necessary to establish more conclusive evidence.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn summary, probiotics offer a multifaceted approach to managing GI symptoms and mood states in bodybuilders through various mechanisms. Given the unique dietary and training challenges faced by bodybuilders, incorporating probiotics into their regimen may provide significant benefits for GI health and mental states, ultimately supporting their performance and recovery. As with any supplement, careful consideration and individualized approaches are essential for achieving the best outcomes.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eGI; Gastrointestinal, GSRS; Gastrointestinal Symptom Rating Scale, BRUMS; Brunel Mood Scale, IPAQ; International Physical Activity Questionnaire, GALT; gut-associated lymphoid tissue, GABA; gamma-aminobutyric acid, RCT; randomized clinical trial\u003c/p\u003e\n"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWritten informed consent was obtained from participants before participation in the research project. The current trial received approval from Medical Ethics Committee of Tehran University of Medical Sciences, Tehran, Iran (IR. TUMS. MEDICNE.REC.1403.157). This study was conducted in accordance with the Declaration of Helsinki.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eNot applicable.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analyzed during this study are included in this published article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe trial funding was supported by Tehran University of Medical Science (Grant number: 72295). The funder is not involved in the study design, collection, management, analysis and interpretation, writing of the manuscript, and the decision to submit the report for publication, including whether they will have ultimate authority over any of these activities.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor\u0026rsquo;s contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNastaran Mahmoudi Shirkoohi: Methodology; writing \u0026ndash; original draft; writing \u0026ndash; review and editing. Alireza Chegol: Writing \u0026ndash; original draft; writing \u0026ndash; review and editing. Hamed Mohammadi: Methodology; software; writing \u0026ndash; original draft; writing \u0026ndash; review and editing. Sakineh Shab-bidar: Methodology; validation; Formal analysis; visualization; writing \u0026ndash; review and editing. Siroos Choobineh: Methodology; writing \u0026ndash; review and editing. Kurosh Djafarian: Conceptualization; methodology; project administration; resources; writing \u0026ndash; review and editing.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe sincerely thank the Tak Gen Zist Pharmaceutical Company for their support.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eMaughan, R.J. and S.M. Shirreffs, \u003cem\u003eIOC consensus conference on nutrition in sport, 25\u0026ndash;27 October 2010, International Olympic Committee, Lausanne, Switzerland.\u003c/em\u003e Journal of sports sciences, 2011. \u003cstrong\u003e29\u003c/strong\u003e(sup1): p. S1-S1.\u003c/li\u003e\n\u003cli\u003eSchoenfeld, B.J., A.A. Aragon, and J.W. 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Khoshouei, \u003cem\u003eComparison between psychometric characteristics of persian version of the gastrointestinal symptoms rating scale in functional gastrointestinal disorders and normal groups.\u003c/em\u003e 2012.\u003c/li\u003e\n\u003cli\u003eFAROKHI, A., E. MOTESHAREIE, and R. ZEIDABADY, \u003cem\u003eValidity and reliability of Persian version of Brunel mood scale 32 items.\u003c/em\u003e 2013.\u003c/li\u003e\n\u003cli\u003eBagherpour, T., H.A. Hashim, and S. Saha, \u003cem\u003eFactorial Validity and Internal Consistency of Iranian Adapted Brunel Mood Scale among Iranian Young Female Taekwondo Athletes.\u003c/em\u003e J Phys Act Sport Exerc, 2013. \u003cstrong\u003e1\u003c/strong\u003e(1): p. 74-77.\u003c/li\u003e\n\u003cli\u003eWang, L., et al., \u003cem\u003eEffects of probiotic supplementation on 12 min run performance, mood management, body composition and gut microbiota in amateur marathon runners: A double-blind controlled trial.\u003c/em\u003e Journal of Exercise Science \u0026amp; Fitness, 2024. \u003cstrong\u003e22\u003c/strong\u003e(4): p. 297-304.\u003c/li\u003e\n\u003cli\u003eRoberts, J.D., et al., \u003cem\u003eAn exploratory investigation of endotoxin levels in novice long distance triathletes, and the effects of a multi-strain probiotic/prebiotic, antioxidant intervention.\u003c/em\u003e Nutrients, 2016. \u003cstrong\u003e8\u003c/strong\u003e(11): p. 733.\u003c/li\u003e\n\u003cli\u003eWest, N.P., et al., \u003cem\u003eLactobacillus fermentum (PCC\u0026reg;) supplementation and gastrointestinal and respiratory-tract illness symptoms: a randomised control trial in athletes.\u003c/em\u003e Nutrition journal, 2011. \u003cstrong\u003e10\u003c/strong\u003e: p. 1-11.\u003c/li\u003e\n\u003cli\u003ePugh, J.N., et al., \u003cem\u003eProbiotic supplementation increases carbohydrate metabolism in trained male cyclists: A randomized, double-blind, placebo-controlled crossover trial.\u003c/em\u003e American Journal of Physiology-Endocrinology and Metabolism, 2020. \u003cstrong\u003e318\u003c/strong\u003e(4): p. E504-E513.\u003c/li\u003e\n\u003cli\u003eNoorifard, M., et al., \u003cem\u003eEffect of probiotic supplementation on oxidative stress enzymes and mental health of athletes.\u003c/em\u003e Annals of Military and Health Sciences Research, 2019. \u003cstrong\u003e17\u003c/strong\u003e(1).\u003c/li\u003e\n\u003cli\u003eSalleh, R.M., et al., \u003cem\u003eEffects of probiotics on anxiety, stress, mood and fitness of badminton players.\u003c/em\u003e Nutrients, 2021. \u003cstrong\u003e13\u003c/strong\u003e(6): p. 1783.\u003c/li\u003e\n\u003cli\u003eCriswell, D.S., et al., \u003cem\u003eChronic exercise and the pro-inflammatory response to endotoxin in the serum and heart.\u003c/em\u003e Immunology letters, 2004. \u003cstrong\u003e95\u003c/strong\u003e(2): p. 213-220.\u003c/li\u003e\n\u003cli\u003eFukui, H., \u003cem\u003eIncreased intestinal permeability and decreased barrier function: does it really influence the risk of inflammation?\u003c/em\u003e Inflammatory intestinal diseases, 2016. \u003cstrong\u003e1\u003c/strong\u003e(3): p. 135-145.\u003c/li\u003e\n\u003cli\u003eCinca-Morros, S. and J. \u0026Aacute;lvarez-Herms, \u003cem\u003eThe Importance of Maintaining and Improving a Healthy Gut Microbiota in Athletes as a Preventive Strategy to Improve Heat Tolerance and Acclimatization.\u003c/em\u003e Microorganisms, 2024. \u003cstrong\u003e12\u003c/strong\u003e(6): p. 1160.\u003c/li\u003e\n\u003cli\u003eLennon, S., \u003cem\u003eThe effects of probiotic supplementation on endurance athletes: Influences on gut dysfunction, microbiome, and gut discomfort\u003c/em\u003e. 2022: The University of Memphis.\u003c/li\u003e\n\u003cli\u003eSmith, K.A., et al., \u003cem\u003eGastrointestinal pathophysiology during endurance exercise: endocrine, microbiome, and nutritional influences.\u003c/em\u003e European Journal of Applied Physiology, 2021. \u003cstrong\u003e121\u003c/strong\u003e(10): p. 2657-2674.\u003c/li\u003e\n\u003cli\u003eM\u0026ouml;rbe, U.M., et al., \u003cem\u003eHuman gut-associated lymphoid tissues (GALT); diversity, structure, and function.\u003c/em\u003e Mucosal immunology, 2021. \u003cstrong\u003e14\u003c/strong\u003e(4): p. 793-802.\u003c/li\u003e\n\u003cli\u003eStarkie, R., et al., \u003cem\u003eCirculating monocytes are not the source of elevations in plasma IL-6 and TNF-\u0026alpha; levels after prolonged running.\u003c/em\u003e American Journal of Physiology-Cell Physiology, 2001. \u003cstrong\u003e280\u003c/strong\u003e(4): p. C769-C774.\u003c/li\u003e\n\u003cli\u003eŁagowska, K. and J. Bajerska, \u003cem\u003eProbiotic supplementation and respiratory infection and immune function in athletes: systematic review and meta-analysis of randomized controlled trials.\u003c/em\u003e Journal of Athletic Training, 2021. \u003cstrong\u003e56\u003c/strong\u003e(11): p. 1213-1223.\u003c/li\u003e\n\u003cli\u003eWosinska, L., et al., \u003cem\u003eThe potential impact of probiotics on the gut microbiome of athletes.\u003c/em\u003e Nutrients, 2019. \u003cstrong\u003e11\u003c/strong\u003e(10): p. 2270.\u003c/li\u003e\n\u003cli\u003eDi Dio, M., et al., \u003cem\u003eEffects of Probiotic supplementation on sports performance and performance-related features in athletes: a systematic review.\u003c/em\u003e International Journal of Environmental Research and Public Health, 2023. \u003cstrong\u003e20\u003c/strong\u003e(3): p. 2226.\u003c/li\u003e\n\u003cli\u003eSivamaruthi, B.S., P. Kesika, and C. Chaiyasut, \u003cem\u003eEffect of probiotics supplementations on health status of athletes.\u003c/em\u003e International Journal of Environmental Research and Public Health, 2019. \u003cstrong\u003e16\u003c/strong\u003e(22): p. 4469.\u003c/li\u003e\n\u003cli\u003eBravo, J.A., et al., \u003cem\u003eIngestion of Lactobacillus strain regulates emotional behavior and central GABA receptor expression in a mouse via the vagus nerve.\u003c/em\u003e Proceedings of the National Academy of Sciences, 2011. \u003cstrong\u003e108\u003c/strong\u003e(38): p. 16050-16055.\u003c/li\u003e\n\u003cli\u003ePinto-Sanchez, M.I., et al., \u003cem\u003eProbiotic Bifidobacterium longum NCC3001 reduces depression scores and alters brain activity: a pilot study in patients with irritable bowel syndrome.\u003c/em\u003e Gastroenterology, 2017. \u003cstrong\u003e153\u003c/strong\u003e(2): p. 448-459. e8.\u003c/li\u003e\n\u003cli\u003eZhang, L., R. Zhang, and L. Li, \u003cem\u003eEffects of probiotic supplementation on exercise and the underlying mechanisms.\u003c/em\u003e Foods, 2023. \u003cstrong\u003e12\u003c/strong\u003e(9): p. 1787.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Table 1","content":"\u003cp\u003eTable 1 is available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-gastroenterology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bmge","sideBox":"Learn more about [BMC Gastroenterology](http://bmcgastroenterol.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/bmge/default.aspx","title":"BMC Gastroenterology","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"probiotic, gastrointestinal symptoms, mood states, bodybuilders","lastPublishedDoi":"10.21203/rs.3.rs-7147381/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7147381/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e\u003cp\u003eEmerging evidence suggests that probiotics may alleviate Gastrointestinal (GI) disturbances and positively influence mood by modulating the gut-brain axis. However, there is limited research specifically examining the effects of probiotic supplementation on GI symptoms and mood in bodybuilders. This study aims to investigate the effect of probiotics on Gastrointestinal health and psychological well-being in this population.\u003c/p\u003e\u003ch2\u003eMethod\u003c/h2\u003e\u003cp\u003eThis study was conducted as a randomized, double-blind, placebo-controlled trial involving 36 unprofessional bodybuilders. Participants were randomly assigned to two groups to receive either 2\u0026nbsp;billion colony-forming units (CFU) of probiotic (n\u0026thinsp;=\u0026thinsp;18) or a placebo (n\u0026thinsp;=\u0026thinsp;18) for 12 weeks. The change in symptoms wase determined using the Gastrointestinal Symptom Rating Scale (GSRS) and the Brunel Mood Scale (BRUMS).\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eAfter 12 weeks of probiotic supplementation, athletes experienced a significant reduction in overall GSRS )p\u0026thinsp;=\u0026thinsp;0.005), reflux (p\u0026thinsp;=\u0026thinsp;0.04), and abdominal pain (p\u0026thinsp;=\u0026thinsp;0.007) scores and significantly improved vigor (p\u0026thinsp;=\u0026thinsp;0.002), peace (p\u0026thinsp;=\u0026thinsp;0.004), and happiness score (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) compared to their baseline measurements. Between-group analysis also revealed a significant reduction in reflux (p\u0026thinsp;=\u0026thinsp;0.01), abdominal pain (P\u0026thinsp;=\u0026thinsp;0.01), and overall GSRS scores (P\u0026thinsp;=\u0026thinsp;0.02). We observed a significant rise in vigor (p\u0026thinsp;=\u0026thinsp;0.02), peace (p\u0026thinsp;=\u0026thinsp;0.01), and happiness score (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eThe results demonstrated that probiotic supplementation has the potential to improve Gastrointestinal symptoms and mood states in athletes. Further studies with adequate power should be performed.\u003c/p\u003e\u003ch2\u003eTrial registration:\u003c/h2\u003e\u003cp\u003eIranian Registry of Clinical Trials IRCT20240424061562N1, Registration date: 2024\u0026ndash;07\u0026ndash;30, Trial Id: 77722.\u003c/p\u003e","manuscriptTitle":"The Effect of Probiotic Supplementation on Gastro-intestinal Symptoms and Mood States in Unprofessional Bodybuilders: A Randomized, Double Blind, Placebo-Controlled Clinical Trial Study","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-24 09:26:07","doi":"10.21203/rs.3.rs-7147381/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-02-06T12:04:59+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-06T10:42:57+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-04T11:06:40+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-08T12:52:07+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-26T13:42:11+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"303206924597157252070480760515881562607","date":"2025-12-20T06:24:28+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"248124228058584936482772144104762413972","date":"2025-12-19T03:38:22+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"163647079621486152194598990144077420991","date":"2025-12-18T11:31:28+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-05T16:39:12+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"69437471645376050388568021762450081983","date":"2025-10-01T12:37:11+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-16T07:20:43+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-08-05T21:36:57+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-08-04T05:29:16+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-08-03T23:24:09+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Gastroenterology","date":"2025-07-31T07:05:08+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-gastroenterology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bmge","sideBox":"Learn more about [BMC Gastroenterology](http://bmcgastroenterol.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/bmge/default.aspx","title":"BMC Gastroenterology","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"ecc23262-2d7c-4032-bff9-ce2d113fcb67","owner":[],"postedDate":"September 24th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-02-17T13:40:16+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-24 09:26:07","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7147381","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7147381","identity":"rs-7147381","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}