Short-term Effects of Oxygenated Nanobubble-Infused Drinks vs Placebo Drinks on Performance and Fatigue Among Endurance Runners: A Crossover Trial

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Arun Maiya, Pragathi N, Anupama Harihar, Ramyashree Kalmadi, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8840472/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 9 You are reading this latest preprint version Abstract Purpose The study aims to check the short-term effects of oxygenated nanobubble-infused drinks on aerobic and anaerobic performance and fatigue among endurance runners. Oxygenated nanobubble beverages have recently gained a lot of attention as an ergogenic aid due to their potential to increase exercise performance and oxygen availability. The primary research question was whether acute consumption of oxygenated nanobubble beverages enhances aerobic performance, anaerobic performance and fatigue resistance compared to a placebo among endurance runners. Methods A randomized, double blind, placebo-controlled crossover trial was conducted by including twenty recreational endurance runners aged 18–30 years. Participants underwent aerobic fitness testing using the Modified Bruce Protocol Test to assess peak oxygen uptake VO₂peak. Anaerobic capacity and Fatigue index were measured using the Wingate Anaerobic Test. Each participant consumed both oxygenated nanobubble-infused drinks and placebo drinks with 48hours of washout period. Performance results were compared between conditions. Results Consumption of oxygenated nanobubble-infused drinks showed an improvement in aerobic performance, with a mean VO₂peak of 68.5 ± 9.48 ml/kg/min. Anaerobic capacity also increased from 540 ± 113 watts to 582 ± 138 watts, while fatigue index decreased from 45.3 ± 10.03% placebo to 41 ± 10.30%, indicating improved resistance to fatigue compared to the placebo drink. Conclusion Oxygenated nanobubble-infused drinks may provide ergogenic benefits by enhancing aerobic and anaerobic performance and reducing fatigue in endurance runners; however, larger trials are needed to validate effectiveness and underlying mechanisms. Oxygenated nanobubble drinks ergogenic aid aerobic performance endurance runners fatigue index anaerobic performance Figures Figure 1 Introduction Athletic performance has driven a continuous innovation in sports, particularly in terms of ergogenic aid. Oxygen plays an important role in endurance performance, and athletes have many ways to enhance oxygen delivery during training or competition( 1 ). Oxygen availability is a key role in aerobic metabolism, which makes it a primary determinant of sustained exercise capacity and an important factor in delaying fatigue, as it controls adenosine triphosphate (ATP) resynthesis and mitochondrial oxidative phosphorylation during prolonged physical activity( 2 ). Several studies have examined the physiological advantages of increased oxygen availability during exercise. These studies found that improved performance is a result of several factors, including increased VO₂max, decreased submaximal heart rate, and blood lactate concentrations( 3 , 4 ). Aerobic performance reflects the combined capacity of circulatory, pulmonary and muscular systems to deliver and consume oxygen during exercise. A key factor in determining performance in endurance athletes, especially long-distance runners, is peak oxygen uptake(VO₂peak), reflecting both central oxygen delivery and peripheral oxygen extraction( 5 ). High-intensity and long-term exercises are known to interfere with oxygen homeostasis; even in normoxic settings, strenuous work has been shown to lower the arterial oxygen saturation (SpO₂). Experimental studies have shown that exercise-induced haemoglobin desaturation and decreases in peripheral SpO₂ occur during cycling and other endurance sports( 6 ). It has been demonstrated that hyperoxic therapies, such as additional oxygen inhalation, decrease the blood lactate accumulation, lower the heart rate and increase exercise tolerance. However, these methods are not feasible to use because it requires such specialized equipment, regulated conditions or face masks9( 7 – 11 ). To overcome these challenges, a new class of dietary supplements, which includes rich oxygen beverages, has emerged. Initial study on traditional oxygenated water, produced by simply dissolving oxygen into a liquid, has failed to demonstrate a performance benefit, mainly due to poor stability of the dissolved oxygen, which tends to dissipate rapidly over time. Currently, there is the development of a novel technology: oxygen nanobubble water (NBO2W). This technology encapsulates oxygen molecules within nanometer-sized bubbles, which are significantly more stable and resistant to buoyancy and gas diffusion ( 12 ). Furthermore, oxygenated nanobubble water enhances hypoxic areas in tumors; when taken orally, it transports the nanobubble structure to intended organs without disintegrating( 13 ). This innovation presents a potentially more effective and practical method for delivering supplemental oxygen to athletes. Therefore, the present study aims to evaluate the short-term effects of oxygenated nanobubble-infused drinks on aerobic performance, assessed as VO₂ peak using the Modified Bruce Protocol Test, anaerobic performance, assessed as anaerobic capacity and fatigue resistance assessed as fatigue index using the Wingate Anaerobic test among healthy recreational endurance runners. Methods Study Design Institutional Research and Ethics Committee approval was obtained (IEC1: 437/2024), and the study was registered with CTRI/2025/03/082772. This was a randomized, placebo-controlled crossover study that was conducted from February 2025 to July 2025. A crossover design was chosen to allow for within-subject comparisons by having each participant serve as their own control. Eligibility criteria Inclusion criteria include young adults aged between 18 to 30 years of both genders, recreational endurance runners and individuals who have participated in at least three institutional-level running events representing institutions were included in the study. Exclusion criteria includes, individuals with history of chronic disease that may limit physical activity, any fluid imbalance disorders or allergies to citrus resulting from oxygenated nanobubble infused soft drinks, presence of systemic vascular or autoimmune disorders affecting fluid balance, any known hypertensive or cardiovascular disorders that may be exacerbated with fluid infusions. Out of the 38 screened patients, 20 met the eligibility criteria and completed the trial. Procedure A total of 38 participants were screened, and 20 were included in this study. The reasons for excluding 18 participants were: ankle sprains ( 5 ), chronic conditions ( 4 ), autoimmune diseases ( 4 ), and recent fractures ( 5 ). After baseline assessment, the participants were randomly assigned to either the placebo group or the nanobubble drink group. The participants consumed a 250 ml of IOTA oxygenated nanobubble-infused drink or placebo drink and immediately performed their assigned tests. After a 48-hour washout period, they were crossed over to receive the other drink, and the remaining test was performed. (1) Baseline assessment Body composition was assessed using bioelectrical impedance analysis (BIA). Skeletal muscle mass, visceral fat level and body fat percentage were recorded( 14 , 15 ). Waist circumference was measured using a tape while the participant stood upright with arms relaxed at their sides, feet together, and abdomen relaxed. A horizontal measurement was taken at the narrowest point of the torso, which is situated above the umbilicus and below the xiphoid process( 16 ). Flexibility was assessed using a sit-to-reach test (for hamstring/lower back flexibility). Participants were asked to sit on the floor with legs fully extended and heels placed against the box. They were instructed to reach forward slowly, without bending the knees, and held the maximal position for two seconds. The best of three attempts was documented( 17 ). 1-Repetition Maximum (1RM) Strength Testing- Maximal strength of the upper and lower limbs was assessed using a body-solid bench press and a leg press machine, respectively. Participants completed a familiarization session before testing. A standard warm-up consisting of several submaximal repetitions of the respective exercise was performed before initiating the assessment. Each participant performed up to four attempts with 3-5minutes of rest between trials. The initial load was set at approximately 50-7-% of the participant’s perceived maximal capacity. Resistance progressively increased by 5–10% for upper limb and 10–20% for lower limb exercises after each successful lift until a failed attempt occurred. The heaviest successfully lifted load was recorded as the absolute 1RM (or multiple RM), and the 1RM value was estimated using Epley’s Eq. (18,19). Muscular endurance was assessed using a field-based push-up test for the upper limb and squat test for the lower limb, performed to volitional fatigue without rest. For push-ups, participants started in the standard prone position with their hands placed under the shoulders, a straight back, and toes as the pivot point. Repetitions were counted only if participants fully extended their elbows and returned to the starting position without allowing the abdomen to contact the mat. The maximum number of continuous repetitions was recorded( 20 ). (2) Cardiopulmonary fitness testing-Modified Bruce treadmill test Cardiopulmonary fitness was evaluated using a treadmill test based on the Modified Bruce Protocol. The procedure was explained to all the participants, and written informed consent was obtained prior to testing. Participants were ensured to be dressed in comfortable exercise clothing and suitable footwear. Baseline physiological parameters, including heart rate (HR), oxygen saturation (SpO₂), blood pressure (BP), and Borg Rating of Perceived Exertion (RPE) were monitored throughout the test. The VO₂ peak was calculated using the ACSM Treadmill Running Equation. The modified Bruce protocol was employed, which begins at a lower speed and incline and progresses with smaller workload increments compared to the standard Bruce protocol. The test comprises multiple 3-minute stages, with both treadmill speed and incline increased at each stage to progressively elevate cardiovascular demand. Heart rate (HR), peripheral oxygen saturation (SPO2) and blood pressure (BP) were measured at the end of each stage. Borg Rating of Perceived Exertion (RPE) was recorded at each stage. Throughout testing, participants were closely monitored for any signs of discomfort, fatigue, or adverse symptoms such as dizziness or chest pain( 21 , 22 ). Termination criteria: Achievement of the target HR (usually 85% of the age-predicted max HR) or the VO₂ max plateau. The participants were asked to stop due to fatigue Development of adverse symptoms (e.g., angina, severe breathlessness, ECG abnormalities, dangerous BP changes) Clinical decision making by supervising examiner Post-Test Procedure: The treadmill was returned to 0% inclined at low speed for active recovery. Monitoring of HR, BP, and SpO₂ was continued immediately and at 1 minute and 3 minutes posttest. The test duration, HR, BP, and RPE are documented, and the VO2 peak is calculated. VO2 peak estimation was performed via the ACSM Treadmill Running Equation: VO2 peak (ml/kg/min) = (0.1 × S) + (1.8 × S × G) + 3.5 where: S = Speed in meters per minute (1 mph ≈ 26.8 m/min) G = treadmill grade as a decimal (e.g., 10% = 0.10) 3.5 = resting oxygen consumption ( 3 ) Wingate anaerobic test The Wingate Anaerobic Test is a short-duration, high-intensity cycling test designed to measure anaerobic power, anaerobic capacity, and the fatigue index. It is one of the most widely used laboratory protocols to assess performance in the ATP-PC and glycolytic energy systems. Equipment requirements: Monark cycle ergometer, calibrated load weights, and stopwatch. The test procedure was explained, and familiarization of the equipment was done to the participants. Participants’ body mass was measured and recorded to determine the resistance load. The cycle seat and handlebars were adjusted to the participant’s comfort. A 5–10 min warm-up with light pedalling and 2–3 short sprints was performed to prepare the muscles for maximal effort. A standard load = 0.075 × body mass (kg) was set and the resistance was applied instantly at the start of the test. The participant begins pedaling at the maximal speed with no resistance for 5–10 seconds. At the command "Go", the predetermined resistance is applied instantly to the flywheel. The participant pedals all-out for 30 seconds against the fixed load. Strong verbal encouragement is given to maintain maximal cadence throughout( 23 , 24 ). Measurements and calculations: Peak power (PP) = highest 5-second power output during the test. Anaerobic capacity (AC) = average power over the full 30 seconds. Fatigue Index (FI) = percentage drop from peak to lowest 5-second power output The peak power and anaerobic capacity were calculated and recorded in watts (W) and watts per kilogram body weight (W/kg − 1); the FI was calculated as a percentage Peak power output = force (kg)* distance (m) ÷ time (s) Distance = number of revolutions during the 5-seconds* distance per revolution (m) Fatigue index = ((peak power – lowest power) ÷ (peak power))* 100 Anaerobic capacity = average peak power output Data analysis Data analysis was performed via Jamovi 2.4.11 software. Descriptive statistics were used to summarize the demographic and baseline characteristics of the participants. Means and standard deviations (mean ± SD) are reported for continuous variables. The Shapiro‒Wilk test was used to test for normality. A paired t test was used to compare the effects of the nanobubble drink and placebo drink within and between the groups. P values less than 0.05 were considered significant between the pre- and post-intervention data. Results A total of twenty participants were included in the study. Baseline characteristics, including age, body mass index (BMI), waist circumference (WC), skeletal muscle mass (SMM), body fat percentage and visceral fat level, were assessed (Table 1 ). All twenty participants completed the study with no adverse events or protocol violations. Table 1 Baseline characteristics of the participants (n = 20) Variable Mean ± SD Age (yrs) 22.6 ± 2.36 BMI (kg/m2) 24.1 ± 4.13 WC (cm) 88.4 ± 8.93 Skeletal muscle mass (kg) 32.1 ± 4.06 Percentage body fat (%) 20.6 ± 8.77 Visceral fat level 5.95 ± 4.21 As shown above, the participants had a healthy body composition, which justifies their selection for this study. Table 2 Musculoskeletal examinations of the participants (n = 20) Variable Mean ± SD Flexibility (cm) 7.67 ± 2.16 UL endurance (repetitions) 25.3 ± 7.5 LL endurance (repetitions) 91.1 ± 44.6 UL 1RM (lbs) 125.8 ± 30.5 LL 1 RM (lbs) 239.2 ± 48.2 The musculoskeletal profile of the endurance runners revealed a disparity between upper and lower body strength and endurance, which is consistent with the adaptations of endurance runners. Table 3 Mean VO₂ peak following the treadmill test: Placebo vs. the Nanobubble drink VO₂ peak (ml/kg/min) Mean ± SD df p value Placebo drink 64.3 ± 7.55 19 0.001 Nanobubble drink 68.5 ± 9.48 Table 3 summarizes the cardiorespiratory fitness outcomes following the Modified Bruce protocol treadmill test. The VO₂ peak is increased by 6.5%. After the consumption of oxygenated nanobubble-infused drink, indicating a positive trend in aerobic performance. Table 4 Mean anaerobic capacity following the Wingate test: Placebo vs nanobubble drink Wingate test: Anaerobic capacity (watts) Mean ± SD df p value Placebo drink 540 ± 113 19 0.009 Nanobubble drink 582 ± 138 Table 4 summarizes the anaerobic capacity, measured through the Wingate anaerobic test, which is increased by 7.7% with consumption of the oxygenated nanobubble-infused drink as compared to placebo drink. Table 5 The mean Fatigue index following the Wingate test: Placebo vs nanobubble drink Wingate test: Fatigue Index (%) Mean ± SD df p value Placebo drink 45.3 ± 10.03 19 < 0.001 Nanobubble drink 41 ± 10.30 Table 5 describes the fatigue index, and it shows that it dropped by 9.5%, indicating better fatigue resistance, reflecting improved resistance to fatigue following oxygenated nanobubble-infused drink consumption. Discussion The present study evaluated the effects of an oxygenated nanobubble-infused drink on both aerobic performance during the modified Bruce protocol treadmill test and anaerobic performance during the Wingate anaerobic test, an aspect that has been mostly overlooked in earlier studies. To the best of our knowledge, this is the first study to report performance enhancement through oxygenated nanobubble-infused drink consumption, demonstrating improvement in both aerobic and anaerobic capacity. This study explored the short-term ergogenic effects of oxygenated nanobubble-infused drinks on aerobic and anaerobic performance as well as fatigue resistance among recreational endurance runners. With a crossover trial design, participants underwent both placebo and nanobubble interventions, allowing for within-subject comparisons. The result showed that drinking a nanobubble drink consistently improved the VO₂ peak, anaerobic capacity, and fatigue index, indicating potential performance benefits from this novel intervention. The result demonstrated that supplementing with oxygen nanobubbles produced quantifiable improvements in these measures when compared to placebo, which aligns with and extends findings from earlier studies on oxygen-rich interventions( 25 ). The participants’ mean age was 22.6 ± 2.37 years, and their BMI was 24.1 ± 4.12 kg/m². The mean waist circumference in this study was 88.4 ± 8.93, which was in the low to high-risk category. Adequate skeletal muscle mass and endurance in this population indicated that they are a moderately trained population. The proportion of body fat percentage and skeletal muscle mass (32.1 ± 4.06 and 20.6 ± 8.77, respectively) in recreational endurance runners may represent adaptations linked to regular endurance activity( 26 ). The average sit-and-reach flexibility score among participants in our study was approximately 7.7cm, which indicates that they have good flexibility levels. According to Mayorga-Vega D et al. (2024), the normative values for adults range from + 6 to + 16 cm. Both male and female individuals have adequate flexibility for endurance performance, as indicated by the average score( 27 ). Upper-body strength in the study participants was relatively low, wity mean 1RM bench press value of 125.8 ± 30.5 lb and strength to bodyweight ratio of 0.79. These values fall below reported normative values for recreationally trained young adults. These findings reflect the sports-specific training profile of endurance runners, who more likely prioritise aerobic capacity and lower limb strength than upper-body strength. Previous literature indicates that endurance athletes often demonstrate lower absolute and relative upper-body strength compared with strength and power trained individuals( 28 ). Lower-body strength, on other hand, was significantly higher (239.2 ± 48.2lb), which is indicative of metabolic and biomechanical demands of endurance runners, they need the repetitive production of force from the lower extremities. This difference in strength between upper and lower body is in line with normal physiological changes which seen in populations that have received endurance training( 29 ). The aerobic capacity (VO₂ peak) of participants improved from 64.3 ± 7.55ml/kg/min (placebo) to 68.5 ± 9.4855ml/kg/min following oxygenated nanobubble-infused drink consumption. Representing an approximate 6.5% increase that may be of physiological significance for endurance athletes, as even slight improvements can influence competition outcomes. The observed increase in VO₂ peak aligns with the idea that an oxygenated nanobubble-infused drink could enhance oxygen supply to working muscles, hence increasing aerobic capacity during high-intensity endurance efforts. This is in line with other studies on ozone nanobubble water in animal models, where the therapy improved mice’s total exercise endurance and oxygen consumption (VO₂)( 30 ). Similarly, the study revealed a statistically significant improvement in anaerobic capacity among recreational endurance runners after consumption of oxygenated nanobubble-infused drink (p = 0.009), and a significant reduction in fatigue index (p = < 0.001) was observed. This indicates better buffering against metabolic byproducts like lactate, potentially delaying the onset of muscular fatigue. This is consistent with results from a randomized, placebo-controlled study by King et al on competitive cyclists that found that an oxygen nanobubble beverage increased repeated sprint peak power by 7.1% and 16.1-km time trial performance by 2.4% due to increased submaximal and high-intensity efforts( 25 ). The concept of increased exercise tolerance and delayed fatigue was supported by another randomized crossover trial that found that single- nanosized oxygen nanobubble water increased work rate at lactate threshold by 5%( 31 ). This study has several limitations. The small sample size (n = 20), which restricts the generalizability of the results and raises the risk of statistical errors, especially when assessing small-to moderate effect sizes. Furthermore, long-term safety, efficacy and any physiological changes linked to repeated use of oxygenated nanobubble-infused drinks are yet unknown because only short-term effects were investigated. Another limitation is the lack of physiological and biochemical indicators, including blood lactate, oxygen saturation and respiratory exchange ratio. Further research using these metrics would offer a more in-depth mechanistic understanding of the physiological effects of oxygenated nanobubble supplementation. Future research should investigate dose-response relationships, long-term supplementation, and underlying physiological mechanisms. Comparative studies with alternative oxygen delivery techniques may further validate the oxygenated nanobubble-infused drink as a non-invasive ergogenic aid. Conclusions Oxygenated nanobubble-infused drinks showed the ability to enhance both aerobic and anaerobic performance metrics in recreational endurance runners. Improvement observed in anaerobic capacity and fatigue resistance aligns with earlier findings on oxygen-rich supplementation, which indicate potential ergogenic benefit. With the growing interest in non-pharmacological performance aids, nanobubble technology-based oxygen supplementation represents a novel and practical tool in the athletic population; however, larger trials are needed to validate effectiveness and underlying mechanisms. Declarations Conflict of interest The authors declare no conflict of interest Ethical approval The study was conducted in accordance with the Declaration of Helsinki, and the Institutional Ethical Committee’s approval was obtained. IEC number: IEC1: 437/2024 Informed consent Written informed consent was obtained from all the participants Funding This study was funded by IOTA Nanotechnology Private Limited Author Contribution All authors contributed equally to the present study. Conceptualization was performed by A.M. Methodology was developed by A.M, B.C and M.S. Formal analysis and investigation were carried out by A.M, P.N, R.K, and M.S. Manuscript draft preparation was completed by P.N, R.M and A.H. Testing and data collection were conducted by P.N and R.K. All authors reviewed, edited, and approved the final manuscript. Acknowledgement We would like to thank all of the participants for their time and effort during the study. We would also like to acknowledge• Department of Physiotherapy, Center for Podiatry & Diabetic Foot Care and Research, and Department of Exercise and Sports Science from Manipal College of Health Professions, Manipal• IOTA Nanotechnology Pvt. Ltd., for supplying oxygenated nanobubbles and placebo drinks and for their valuable input throughout the project.We would also like to acknowledge the guidance, encouragement, and constructive feedback provided by our colleagues and research assistants, whose expertise greatly enhanced the quality of this work. References Hirayama H, Yamamoto M. Effects of Canoe Ergometer Training in Normal and Hypoxic Environment in Elite Canadian Canoeists. Journal of Training Science for Exercise and Sport. 2011;23(1):63–75. Ross R, Arena R, Myers J, Kokkinos P, Kaminsky LA. Update to the 2016 American Heart Association cardiorespiratory fitness statement. Prog Cardiovasc Dis. 2024;83:10–5. Piantadosi CA. “Oxygenated” water and athletic performance. 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Positive effects of single-nanosized oxygen nanobubble water on exercise tolerance: A randomized crossover trial. Current Research in Physiology. 2025;8:100159. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Revision requested 30 Apr, 2026 Reviews received at journal 27 Apr, 2026 Reviews received at journal 19 Mar, 2026 Reviewers agreed at journal 17 Mar, 2026 Reviewers agreed at journal 14 Mar, 2026 Reviewers invited by journal 13 Mar, 2026 Editor assigned by journal 11 Feb, 2026 Submission checks completed at journal 11 Feb, 2026 First submitted to journal 10 Feb, 2026 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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Arun Maiya","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/ElEQVRIiWNgGAWjYBADHgYG5gNAWoKBgR3CJUYLWwJECzORWkDKDCA0IS38s8+YPWDMsZMxOH7m64afOyzy+JsZGB+8bcOtReJcjrkB47ZkHoMzudtu9p6RKJY4zMBsOBePFoYzPGYSjNuYeQwO5G67wdsmkdhwmIFNmhePFnmIlnoeg/Nvnt38C9Qy/zAD+298WgwgWg7zGNzIYbsNsmUD0BZmfFoMz7CVSSRuO84jeeOZ2W3ZNoliw8OMzZJzzuHWIneGeZvEx23V9nznk5/dfNtWlyd3vPnghzdleLwPAglArHAAzmZsIKAeCuQbENpHwSgYBaNgFKAAAGHdTq+zC5msAAAAAElFTkSuQmCC","orcid":"","institution":"Manipal Academy of Higher Education","correspondingAuthor":true,"prefix":"Dr.","firstName":"G.","middleName":"Arun","lastName":"Maiya","suffix":""},{"id":606209093,"identity":"d210fee0-d794-46e9-850c-e0930b415b8a","order_by":1,"name":"Pragathi N","email":"","orcid":"","institution":"Manipal Academy of Higher Education","correspondingAuthor":false,"prefix":"","firstName":"Pragathi","middleName":"","lastName":"N","suffix":""},{"id":606209095,"identity":"fddaec22-8d13-4eae-8fdc-e02da4cfc73c","order_by":2,"name":"Anupama Harihar","email":"","orcid":"","institution":"Manipal Academy of Higher Education","correspondingAuthor":false,"prefix":"","firstName":"Anupama","middleName":"","lastName":"Harihar","suffix":""},{"id":606209096,"identity":"78443c60-951b-4166-9e51-36cf6b718141","order_by":3,"name":"Ramyashree Kalmadi","email":"","orcid":"","institution":"Manipal Academy of Higher Education","correspondingAuthor":false,"prefix":"","firstName":"Ramyashree","middleName":"","lastName":"Kalmadi","suffix":""},{"id":606209098,"identity":"2482d62b-ba11-44c3-9126-2a6312b9dd12","order_by":4,"name":"Dr. Mukesh Sinha","email":"","orcid":"","institution":"Manipal Academy of Higher Education","correspondingAuthor":false,"prefix":"Dr.","firstName":"Mukesh","middleName":"","lastName":"Sinha","suffix":""},{"id":606209099,"identity":"058fc7d8-78ef-4d6c-a80b-15d673aa616d","order_by":5,"name":"Dr. Bhaskaran Chandrashekaran","email":"","orcid":"","institution":"Manipal Academy of Higher Education","correspondingAuthor":false,"prefix":"Dr.","firstName":"Bhaskaran","middleName":"","lastName":"Chandrashekaran","suffix":""}],"badges":[],"createdAt":"2026-02-10 11:43:03","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8840472/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8840472/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":104702605,"identity":"0068e3e1-fba9-4abe-8176-a3167fd4c7ac","added_by":"auto","created_at":"2026-03-16 08:44:07","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":46388,"visible":true,"origin":"","legend":"\u003cp\u003eBioelectrical Impedance Analysis (BIA)\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-8840472/v1/e7f720be92071a4c912a1d9e.png"},{"id":104702673,"identity":"05fd4116-d2c3-430e-9044-4222aad26264","added_by":"auto","created_at":"2026-03-16 08:44:15","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":679316,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8840472/v1/b8d6fa4c-4d44-434c-9bff-0fc7a71770fe.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Short-term Effects of Oxygenated Nanobubble-Infused Drinks vs Placebo Drinks on Performance and Fatigue Among Endurance Runners: A Crossover Trial","fulltext":[{"header":"Introduction","content":"\u003cp\u003eAthletic performance has driven a continuous innovation in sports, particularly in terms of ergogenic aid. Oxygen plays an important role in endurance performance, and athletes have many ways to enhance oxygen delivery during training or competition(\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e). Oxygen availability is a key role in aerobic metabolism, which makes it a primary determinant of sustained exercise capacity and an important factor in delaying fatigue, as it controls adenosine triphosphate (ATP) resynthesis and mitochondrial oxidative phosphorylation during prolonged physical activity(\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e). Several studies have examined the physiological advantages of increased oxygen availability during exercise. These studies found that improved performance is a result of several factors, including increased VO₂max, decreased submaximal heart rate, and blood lactate concentrations(\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e). Aerobic performance reflects the combined capacity of circulatory, pulmonary and muscular systems to deliver and consume oxygen during exercise. A key factor in determining performance in endurance athletes, especially long-distance runners, is peak oxygen uptake(VO₂peak), reflecting both central oxygen delivery and peripheral oxygen extraction(\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eHigh-intensity and long-term exercises are known to interfere with oxygen homeostasis; even in normoxic settings, strenuous work has been shown to lower the arterial oxygen saturation (SpO₂). Experimental studies have shown that exercise-induced haemoglobin desaturation and decreases in peripheral SpO₂ occur during cycling and other endurance sports(\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e). It has been demonstrated that hyperoxic therapies, such as additional oxygen inhalation, decrease the blood lactate accumulation, lower the heart rate and increase exercise tolerance. However, these methods are not feasible to use because it requires such specialized equipment, regulated conditions or face masks9(\u003cspan additionalcitationids=\"CR8 CR9 CR10\" citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eTo overcome these challenges, a new class of dietary supplements, which includes rich oxygen beverages, has emerged. Initial study on traditional oxygenated water, produced by simply dissolving oxygen into a liquid, has failed to demonstrate a performance benefit, mainly due to poor stability of the dissolved oxygen, which tends to dissipate rapidly over time. Currently, there is the development of a novel technology: oxygen nanobubble water (NBO2W). This technology encapsulates oxygen molecules within nanometer-sized bubbles, which are significantly more stable and resistant to buoyancy and gas diffusion (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e). Furthermore, oxygenated nanobubble water enhances hypoxic areas in tumors; when taken orally, it transports the nanobubble structure to intended organs without disintegrating(\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e). This innovation presents a potentially more effective and practical method for delivering supplemental oxygen to athletes.\u003c/p\u003e \u003cp\u003eTherefore, the present study aims to evaluate the short-term effects of oxygenated nanobubble-infused drinks on aerobic performance, assessed as VO₂ peak using the Modified Bruce Protocol Test, anaerobic performance, assessed as anaerobic capacity and fatigue resistance assessed as fatigue index using the Wingate Anaerobic test among healthy recreational endurance runners.\u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003eStudy Design\u003c/h2\u003e\n \u003cp\u003eInstitutional Research and Ethics Committee approval was obtained (IEC1: 437/2024), and the study was registered with CTRI/2025/03/082772. This was a randomized, placebo-controlled crossover study that was conducted from February 2025 to July 2025. A crossover design was chosen to allow for within-subject comparisons by having each participant serve as their own control.\u003c/p\u003e\n\u003c/div\u003e\n\u003ch3\u003eEligibility criteria\u003c/h3\u003e\n\u003cp\u003eInclusion criteria include young adults aged between 18 to 30 years of both genders, recreational endurance runners and individuals who have participated in at least three institutional-level running events representing institutions were included in the study. Exclusion criteria includes, individuals with history of chronic disease that may limit physical activity, any fluid imbalance disorders or allergies to citrus resulting from oxygenated nanobubble infused soft drinks, presence of systemic vascular or autoimmune disorders affecting fluid balance, any known hypertensive or cardiovascular disorders that may be exacerbated with fluid infusions. Out of the 38 screened patients, 20 met the eligibility criteria and completed the trial.\u003c/p\u003e\n\u003ch3\u003eProcedure\u003c/h3\u003e\n\u003cp\u003eA total of 38 participants were screened, and 20 were included in this study. The reasons for excluding 18 participants were: ankle sprains (\u003cspan class=\"CitationRef\"\u003e5\u003c/span\u003e), chronic conditions (\u003cspan class=\"CitationRef\"\u003e4\u003c/span\u003e), autoimmune diseases (\u003cspan class=\"CitationRef\"\u003e4\u003c/span\u003e), and recent fractures (\u003cspan class=\"CitationRef\"\u003e5\u003c/span\u003e). After baseline assessment, the participants were randomly assigned to either the placebo group or the nanobubble drink group. The participants consumed a 250 ml of IOTA oxygenated nanobubble-infused drink or placebo drink and immediately performed their assigned tests. After a 48-hour washout period, they were crossed over to receive the other drink, and the remaining test was performed.\u003c/p\u003e\n\u003ch3\u003e(1) Baseline assessment\u003c/h3\u003e\n\u003cp\u003eBody composition was assessed using bioelectrical impedance analysis (BIA). Skeletal muscle mass, visceral fat level and body fat percentage were recorded(\u003cspan class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan class=\"CitationRef\"\u003e15\u003c/span\u003e). Waist circumference was measured using a tape while the participant stood upright with arms relaxed at their sides, feet together, and abdomen relaxed. A horizontal measurement was taken at the narrowest point of the torso, which is situated above the umbilicus and below the xiphoid process(\u003cspan class=\"CitationRef\"\u003e16\u003c/span\u003e). Flexibility was assessed using a sit-to-reach test (for hamstring/lower back flexibility). Participants were asked to sit on the floor with legs fully extended and heels placed against the box. They were instructed to reach forward slowly, without bending the knees, and held the maximal position for two seconds. The best of three attempts was documented(\u003cspan class=\"CitationRef\"\u003e17\u003c/span\u003e). 1-Repetition Maximum (1RM) Strength Testing- Maximal strength of the upper and lower limbs was assessed using a body-solid bench press and a leg press machine, respectively. Participants completed a familiarization session before testing. A standard warm-up consisting of several submaximal repetitions of the respective exercise was performed before initiating the assessment. Each participant performed up to four attempts with 3-5minutes of rest between trials. The initial load was set at approximately 50-7-% of the participant\u0026rsquo;s perceived maximal capacity. Resistance progressively increased by 5\u0026ndash;10% for upper limb and 10\u0026ndash;20% for lower limb exercises after each successful lift until a failed attempt occurred. The heaviest successfully lifted load was recorded as the absolute 1RM (or multiple RM), and the 1RM value was estimated using Epley\u0026rsquo;s Eq.\u0026nbsp;(18,19). Muscular endurance was assessed using a field-based push-up test for the upper limb and squat test for the lower limb, performed to volitional fatigue without rest. For push-ups, participants started in the standard prone position with their hands placed under the shoulders, a straight back, and toes as the pivot point. Repetitions were counted only if participants fully extended their elbows and returned to the starting position without allowing the abdomen to contact the mat. The maximum number of continuous repetitions was recorded(\u003cspan class=\"CitationRef\"\u003e20\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003e(2) Cardiopulmonary fitness testing-Modified Bruce treadmill test\u003c/h3\u003e\n\u003cp\u003eCardiopulmonary fitness was evaluated using a treadmill test based on the Modified Bruce Protocol. The procedure was explained to all the participants, and written informed consent was obtained prior to testing. Participants were ensured to be dressed in comfortable exercise clothing and suitable footwear. Baseline physiological parameters, including heart rate (HR), oxygen saturation (SpO₂), blood pressure (BP), and Borg Rating of Perceived Exertion (RPE) were monitored throughout the test. The VO₂ peak was calculated using the ACSM Treadmill Running Equation. The modified Bruce protocol was employed, which begins at a lower speed and incline and progresses with smaller workload increments compared to the standard Bruce protocol. The test comprises multiple 3-minute stages, with both treadmill speed and incline increased at each stage to progressively elevate cardiovascular demand. Heart rate (HR), peripheral oxygen saturation (SPO2) and blood pressure (BP) were measured at the end of each stage. Borg Rating of Perceived Exertion (RPE) was recorded at each stage. Throughout testing, participants were closely monitored for any signs of discomfort, fatigue, or adverse symptoms such as dizziness or chest pain(\u003cspan class=\"CitationRef\"\u003e21\u003c/span\u003e, \u003cspan class=\"CitationRef\"\u003e22\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eTermination criteria:\u003c/p\u003e\n\u003cul\u003e\n \u003cli\u003e\n \u003cp\u003eAchievement of the target HR (usually 85% of the age-predicted max HR) or the VO₂ max plateau.\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003eThe participants were asked to stop due to fatigue\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003eDevelopment of adverse symptoms (e.g., angina, severe breathlessness, ECG abnormalities, dangerous BP changes)\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003eClinical decision making by supervising examiner\u003c/p\u003e\n \u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003ePost-Test Procedure:\u003c/p\u003e\n\u003cp\u003eThe treadmill was returned to 0% inclined at low speed for active recovery. Monitoring of HR, BP, and SpO₂ was continued immediately and at 1 minute and 3 minutes posttest. The test duration, HR, BP, and RPE are documented, and the VO2 peak is calculated.\u003c/p\u003e\n\u003cp\u003eVO2 peak estimation was performed via the ACSM Treadmill Running Equation:\u003c/p\u003e\n\u003cp\u003eVO2 peak (ml/kg/min) = (0.1 \u0026times; S) + (1.8 \u0026times; S \u0026times; G)\u0026thinsp;+\u0026thinsp;3.5\u003c/p\u003e\n\u003cp\u003ewhere:\u003c/p\u003e\n\u003cul\u003e\n \u003cli\u003e\n \u003cp\u003eS\u0026thinsp;=\u0026thinsp;Speed in meters per minute (1 mph\u0026thinsp;\u0026asymp;\u0026thinsp;26.8 m/min)\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003eG\u0026thinsp;=\u0026thinsp;treadmill grade as a decimal (e.g., 10% = 0.10)\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003e3.5\u0026thinsp;=\u0026thinsp;resting oxygen consumption\u003c/p\u003e\n \u003c/li\u003e\n\u003c/ul\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n \u003ch2\u003e(\u003cspan class=\"CitationRef\"\u003e3\u003c/span\u003e) Wingate anaerobic test\u003c/h2\u003e\n \u003cp\u003eThe Wingate Anaerobic Test is a short-duration, high-intensity cycling test designed to measure anaerobic power, anaerobic capacity, and the fatigue index. It is one of the most widely used laboratory protocols to assess performance in the ATP-PC and glycolytic energy systems. Equipment requirements: Monark cycle ergometer, calibrated load weights, and stopwatch.\u003c/p\u003e\n \u003cp\u003eThe test procedure was explained, and familiarization of the equipment was done to the participants. Participants\u0026rsquo; body mass was measured and recorded to determine the resistance load. The cycle seat and handlebars were adjusted to the participant\u0026rsquo;s comfort. A 5\u0026ndash;10 min warm-up with light pedalling and 2\u0026ndash;3 short sprints was performed to prepare the muscles for maximal effort. A standard load\u0026thinsp;=\u0026thinsp;0.075 \u0026times; body mass (kg) was set and the resistance was applied instantly at the start of the test. The participant begins pedaling at the maximal speed with no resistance for 5\u0026ndash;10 seconds. At the command \u0026quot;Go\u0026quot;, the predetermined resistance is applied instantly to the flywheel. The participant pedals all-out for 30 seconds against the fixed load. Strong verbal encouragement is given to maintain maximal cadence throughout(\u003cspan class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan class=\"CitationRef\"\u003e24\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eMeasurements and calculations:\u003c/p\u003e\n \u003cp\u003ePeak power (PP) = highest 5-second power output during the test.\u003c/p\u003e\n \u003cp\u003eAnaerobic capacity (AC) = average power over the full 30 seconds.\u003c/p\u003e\n \u003cp\u003eFatigue Index (FI) = percentage drop from peak to lowest 5-second power output\u003c/p\u003e\n \u003cp\u003eThe peak power and anaerobic capacity were calculated and recorded in watts (W) and watts per kilogram body weight (W/kg\u0026thinsp;\u0026minus;\u0026thinsp;1); the FI was calculated as a percentage\u003c/p\u003e\n \u003cul\u003e\n \u003cli\u003e\n \u003cp\u003ePeak power output\u0026thinsp;=\u0026thinsp;force (kg)* distance (m) \u0026divide; time (s)\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003eDistance\u0026thinsp;=\u0026thinsp;number of revolutions during the 5-seconds* distance per revolution (m)\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003eFatigue index = ((peak power \u0026ndash; lowest power) \u0026divide; (peak power))* 100\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003eAnaerobic capacity\u0026thinsp;=\u0026thinsp;average peak power output\u003c/p\u003e\n \u003c/li\u003e\n \u003c/ul\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\n \u003ch2\u003eData analysis\u003c/h2\u003e\n \u003cp\u003eData analysis was performed via Jamovi 2.4.11 software. Descriptive statistics were used to summarize the demographic and baseline characteristics of the participants. Means and standard deviations (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD) are reported for continuous variables. The Shapiro‒Wilk test was used to test for normality. A paired t test was used to compare the effects of the nanobubble drink and placebo drink within and between the groups. P values less than 0.05 were considered significant between the pre- and post-intervention data.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eA total of twenty participants were included in the study. Baseline characteristics, including age, body mass index (BMI), waist circumference (WC), skeletal muscle mass (SMM), body fat percentage and visceral fat level, were assessed (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). All twenty participants completed the study with no adverse events or protocol violations.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eBaseline characteristics of the participants (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVariable\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAge (yrs)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e22.6\u0026thinsp;\u0026plusmn;\u0026thinsp;2.36\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBMI (kg/m2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e24.1\u0026thinsp;\u0026plusmn;\u0026thinsp;4.13\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWC (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e88.4\u0026thinsp;\u0026plusmn;\u0026thinsp;8.93\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSkeletal muscle mass (kg)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e32.1\u0026thinsp;\u0026plusmn;\u0026thinsp;4.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePercentage body fat (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e20.6\u0026thinsp;\u0026plusmn;\u0026thinsp;8.77\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVisceral fat level\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e5.95\u0026thinsp;\u0026plusmn;\u0026thinsp;4.21\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eAs shown above, the participants had a healthy body composition, which justifies their selection for this study.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMusculoskeletal examinations of the participants (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVariable\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFlexibility (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e7.67\u0026thinsp;\u0026plusmn;\u0026thinsp;2.16\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eUL endurance (repetitions)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e25.3\u0026thinsp;\u0026plusmn;\u0026thinsp;7.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLL endurance (repetitions)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e91.1\u0026thinsp;\u0026plusmn;\u0026thinsp;44.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eUL 1RM (lbs)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e125.8\u0026thinsp;\u0026plusmn;\u0026thinsp;30.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLL 1 RM (lbs)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e239.2\u0026thinsp;\u0026plusmn;\u0026thinsp;48.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe musculoskeletal profile of the endurance runners revealed a disparity between upper and lower body strength and endurance, which is consistent with the adaptations of endurance runners.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMean VO₂ peak following the treadmill test: Placebo vs. the Nanobubble drink\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVO₂ peak (ml/kg/min)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003edf\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePlacebo drink\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e64.3\u0026thinsp;\u0026plusmn;\u0026thinsp;7.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNanobubble drink\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e68.5\u0026thinsp;\u0026plusmn;\u0026thinsp;9.48\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e summarizes the cardiorespiratory fitness outcomes following the Modified Bruce protocol treadmill test. The VO₂ peak is increased by 6.5%. After the consumption of oxygenated nanobubble-infused drink, indicating a positive trend in aerobic performance.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMean anaerobic capacity following the Wingate test: Placebo vs nanobubble drink\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWingate test: Anaerobic capacity (watts)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003edf\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePlacebo drink\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e540\u0026thinsp;\u0026plusmn;\u0026thinsp;113\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.009\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNanobubble drink\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e582\u0026thinsp;\u0026plusmn;\u0026thinsp;138\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e summarizes the anaerobic capacity, measured through the Wingate anaerobic test, which is increased by 7.7% with consumption of the oxygenated nanobubble-infused drink as compared to placebo drink.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe mean Fatigue index following the Wingate test: Placebo vs nanobubble drink\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWingate test:\u003c/p\u003e \u003cp\u003eFatigue Index (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003edf\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePlacebo drink\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e45.3\u0026thinsp;\u0026plusmn;\u0026thinsp;10.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNanobubble drink\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e41\u0026thinsp;\u0026plusmn;\u0026thinsp;10.30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e describes the fatigue index, and it shows that it dropped by 9.5%, indicating better fatigue resistance, reflecting improved resistance to fatigue following oxygenated nanobubble-infused drink consumption.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe present study evaluated the effects of an oxygenated nanobubble-infused drink on both aerobic performance during the modified Bruce protocol treadmill test and anaerobic performance during the Wingate anaerobic test, an aspect that has been mostly overlooked in earlier studies. To the best of our knowledge, this is the first study to report performance enhancement through oxygenated nanobubble-infused drink consumption, demonstrating improvement in both aerobic and anaerobic capacity. This study explored the short-term ergogenic effects of oxygenated nanobubble-infused drinks on aerobic and anaerobic performance as well as fatigue resistance among recreational endurance runners. With a crossover trial design, participants underwent both placebo and nanobubble interventions, allowing for within-subject comparisons. The result showed that drinking a nanobubble drink consistently improved the VO₂ peak, anaerobic capacity, and fatigue index, indicating potential performance benefits from this novel intervention. The result demonstrated that supplementing with oxygen nanobubbles produced quantifiable improvements in these measures when compared to placebo, which aligns with and extends findings from earlier studies on oxygen-rich interventions(\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe participants\u0026rsquo; mean age was 22.6\u0026thinsp;\u0026plusmn;\u0026thinsp;2.37 years, and their BMI was 24.1\u0026thinsp;\u0026plusmn;\u0026thinsp;4.12 kg/m\u0026sup2;. The mean waist circumference in this study was 88.4\u0026thinsp;\u0026plusmn;\u0026thinsp;8.93, which was in the low to high-risk category. Adequate skeletal muscle mass and endurance in this population indicated that they are a moderately trained population. The proportion of body fat percentage and skeletal muscle mass (32.1\u0026thinsp;\u0026plusmn;\u0026thinsp;4.06 and 20.6\u0026thinsp;\u0026plusmn;\u0026thinsp;8.77, respectively) in recreational endurance runners may represent adaptations linked to regular endurance activity(\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e). The average sit-and-reach flexibility score among participants in our study was approximately 7.7cm, which indicates that they have good flexibility levels. According to Mayorga-Vega D et al. (2024), the normative values for adults range from +\u0026thinsp;6 to +\u0026thinsp;16 cm. Both male and female individuals have adequate flexibility for endurance performance, as indicated by the average score(\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e). Upper-body strength in the study participants was relatively low, wity mean 1RM bench press value of 125.8\u0026thinsp;\u0026plusmn;\u0026thinsp;30.5 lb and strength to bodyweight ratio of 0.79. These values fall below reported normative values for recreationally trained young adults. These findings reflect the sports-specific training profile of endurance runners, who more likely prioritise aerobic capacity and lower limb strength than upper-body strength. Previous literature indicates that endurance athletes often demonstrate lower absolute and relative upper-body strength compared with strength and power trained individuals(\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e). Lower-body strength, on other hand, was significantly higher (239.2\u0026thinsp;\u0026plusmn;\u0026thinsp;48.2lb), which is indicative of metabolic and biomechanical demands of endurance runners, they need the repetitive production of force from the lower extremities. This difference in strength between upper and lower body is in line with normal physiological changes which seen in populations that have received endurance training(\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe aerobic capacity (VO₂ peak) of participants improved from 64.3\u0026thinsp;\u0026plusmn;\u0026thinsp;7.55ml/kg/min (placebo) to 68.5\u0026thinsp;\u0026plusmn;\u0026thinsp;9.4855ml/kg/min following oxygenated nanobubble-infused drink consumption. Representing an approximate 6.5% increase that may be of physiological significance for endurance athletes, as even slight improvements can influence competition outcomes. The observed increase in VO₂ peak aligns with the idea that an oxygenated nanobubble-infused drink could enhance oxygen supply to working muscles, hence increasing aerobic capacity during high-intensity endurance efforts. This is in line with other studies on ozone nanobubble water in animal models, where the therapy improved mice\u0026rsquo;s total exercise endurance and oxygen consumption (VO₂)(\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSimilarly, the study revealed a statistically significant improvement in anaerobic capacity among recreational endurance runners after consumption of oxygenated nanobubble-infused drink (p\u0026thinsp;=\u0026thinsp;0.009), and a significant reduction in fatigue index (p\u0026thinsp;=\u0026thinsp;\u0026lt;\u0026thinsp;0.001) was observed. This indicates better buffering against metabolic byproducts like lactate, potentially delaying the onset of muscular fatigue. This is consistent with results from a randomized, placebo-controlled study by King et al on competitive cyclists that found that an oxygen nanobubble beverage increased repeated sprint peak power by 7.1% and 16.1-km time trial performance by 2.4% due to increased submaximal and high-intensity efforts(\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e). The concept of increased exercise tolerance and delayed fatigue was supported by another randomized crossover trial that found that single- nanosized oxygen nanobubble water increased work rate at lactate threshold by 5%(\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThis study has several limitations. The small sample size (n\u0026thinsp;=\u0026thinsp;20), which restricts the generalizability of the results and raises the risk of statistical errors, especially when assessing small-to moderate effect sizes. Furthermore, long-term safety, efficacy and any physiological changes linked to repeated use of oxygenated nanobubble-infused drinks are yet unknown because only short-term effects were investigated. Another limitation is the lack of physiological and biochemical indicators, including blood lactate, oxygen saturation and respiratory exchange ratio. Further research using these metrics would offer a more in-depth mechanistic understanding of the physiological effects of oxygenated nanobubble supplementation.\u003c/p\u003e \u003cp\u003eFuture research should investigate dose-response relationships, long-term supplementation, and underlying physiological mechanisms. Comparative studies with alternative oxygen delivery techniques may further validate the oxygenated nanobubble-infused drink as a non-invasive ergogenic aid.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eOxygenated nanobubble-infused drinks showed the ability to enhance both aerobic and anaerobic performance metrics in recreational endurance runners. Improvement observed in anaerobic capacity and fatigue resistance aligns with earlier findings on oxygen-rich supplementation, which indicate potential ergogenic benefit. With the growing interest in non-pharmacological performance aids, nanobubble technology-based oxygen supplementation represents a novel and practical tool in the athletic population; however, larger trials are needed to validate effectiveness and underlying mechanisms.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eConflict of interest\u003c/h2\u003e \u003cp\u003eThe authors declare no conflict of interest\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eEthical approval\u003c/strong\u003e \u003cp\u003e The study was conducted in accordance with the Declaration of Helsinki, and the Institutional Ethical Committee\u0026rsquo;s approval was obtained. IEC number: IEC1: 437/2024\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eInformed consent\u003c/strong\u003e \u003cp\u003e Written informed consent was obtained from all the participants\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThis study was funded by IOTA Nanotechnology Private Limited\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eAll authors contributed equally to the present study. Conceptualization was performed by A.M. Methodology was developed by A.M, B.C and M.S. Formal analysis and investigation were carried out by A.M, P.N, R.K, and M.S. Manuscript draft preparation was completed by P.N, R.M and A.H. Testing and data collection were conducted by P.N and R.K. All authors reviewed, edited, and approved the final manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eWe would like to thank all of the participants for their time and effort during the study. We would also like to acknowledge\u0026bull; Department of Physiotherapy, Center for Podiatry \u0026amp; Diabetic Foot Care and Research, and Department of Exercise and Sports Science from Manipal College of Health Professions, Manipal\u0026bull; IOTA Nanotechnology Pvt. Ltd., for supplying oxygenated nanobubbles and placebo drinks and for their valuable input throughout the project.We would also like to acknowledge the guidance, encouragement, and constructive feedback provided by our colleagues and research assistants, whose expertise greatly enhanced the quality of this work.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eHirayama H, Yamamoto M. Effects of Canoe Ergometer Training in Normal and Hypoxic Environment in Elite Canadian Canoeists. Journal of Training Science for Exercise and Sport. 2011;23(1):63\u0026ndash;75.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRoss R, Arena R, Myers J, Kokkinos P, Kaminsky LA. 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Current Research in Physiology. 2025;8:100159.\u003c/span\u003e\u003c/li\u003e\u003c/ol\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":"sport-sciences-for-health","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ssfh","sideBox":"Learn more about [Sport Sciences for Health](http://link.springer.com/journal/11332)","snPcode":"11332","submissionUrl":"https://submission.nature.com/new-submission/11332/3","title":"Sport Sciences for Health","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Oxygenated nanobubble drinks, ergogenic aid, aerobic performance, endurance runners, fatigue index, anaerobic performance","lastPublishedDoi":"10.21203/rs.3.rs-8840472/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8840472/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003ePurpose\u003c/h2\u003e \u003cp\u003eThe study aims to check the short-term effects of oxygenated nanobubble-infused drinks on aerobic and anaerobic performance and fatigue among endurance runners. Oxygenated nanobubble beverages have recently gained a lot of attention as an ergogenic aid due to their potential to increase exercise performance and oxygen availability. The primary research question was whether acute consumption of oxygenated nanobubble beverages enhances aerobic performance, anaerobic performance and fatigue resistance compared to a placebo among endurance runners.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eA randomized, double blind, placebo-controlled crossover trial was conducted by including twenty recreational endurance runners aged 18\u0026ndash;30 years. Participants underwent aerobic fitness testing using the Modified Bruce Protocol Test to assess peak oxygen uptake VO₂peak. Anaerobic capacity and Fatigue index were measured using the Wingate Anaerobic Test. Each participant consumed both oxygenated nanobubble-infused drinks and placebo drinks with 48hours of washout period. Performance results were compared between conditions.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eConsumption of oxygenated nanobubble-infused drinks showed an improvement in aerobic performance, with a mean VO₂peak of 68.5\u0026thinsp;\u0026plusmn;\u0026thinsp;9.48 ml/kg/min. Anaerobic capacity also increased from 540\u0026thinsp;\u0026plusmn;\u0026thinsp;113 watts to 582\u0026thinsp;\u0026plusmn;\u0026thinsp;138 watts, while fatigue index decreased from 45.3\u0026thinsp;\u0026plusmn;\u0026thinsp;10.03% placebo to 41\u0026thinsp;\u0026plusmn;\u0026thinsp;10.30%, indicating improved resistance to fatigue compared to the placebo drink.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eOxygenated nanobubble-infused drinks may provide ergogenic benefits by enhancing aerobic and anaerobic performance and reducing fatigue in endurance runners; however, larger trials are needed to validate effectiveness and underlying mechanisms.\u003c/p\u003e","manuscriptTitle":"Short-term Effects of Oxygenated Nanobubble-Infused Drinks vs Placebo Drinks on Performance and Fatigue Among Endurance Runners: A Crossover Trial","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-03-16 08:41:00","doi":"10.21203/rs.3.rs-8840472/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-04-30T09:21:41+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-27T20:21:45+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-03-19T19:31:59+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"106596585607832033296253937048528752645","date":"2026-03-17T13:57:52+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"112480333484723332130259708492229770869","date":"2026-03-14T21:40:19+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-03-13T13:10:31+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-02-12T02:28:45+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-02-12T02:27:24+00:00","index":"","fulltext":""},{"type":"submitted","content":"Sport Sciences for Health","date":"2026-02-10T10:47:26+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"sport-sciences-for-health","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ssfh","sideBox":"Learn more about [Sport Sciences for Health](http://link.springer.com/journal/11332)","snPcode":"11332","submissionUrl":"https://submission.nature.com/new-submission/11332/3","title":"Sport Sciences for Health","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"e11e2a70-400a-4f11-9031-37030396f3a9","owner":[],"postedDate":"March 16th, 2026","published":true,"recentEditorialEvents":[{"type":"decision","content":"Revision requested","date":"2026-04-30T09:21:41+00:00","index":"","fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[],"tags":[],"updatedAt":"2026-04-30T09:25:35+00:00","versionOfRecord":[],"versionCreatedAt":"2026-03-16 08:41:00","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8840472","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8840472","identity":"rs-8840472","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}