Dual-Action Grouper Bone and Wakame Hydrolysates Supplement Enhances Exercise Performance and Modulates Gut Microbiota in Mice

Abstract

Background: Sustainable, dual-action ergogenic strategies are underexplored; most products target a single pathway and rarely upcycle seafood sidestreams. We therefore tested an upcycled formulation combining grouper bone hydrolysate and Undaria pinnatifida extract (GU) for ergogenic and microbiota effects in mice. We tested the ergogenic and microbiota modulating effects of GU in mice versus a vehicle and a BCAA control. Methods: GU was prepared via enzymatic hydrolysis of marine by-products and administered to male ICR mice for 4 weeks. Mice were divided into five groups (n = 7/group), receiving a vehicle control, a branched-chain amino acid (BCAA) supplement, or GU at three dose levels (1X, 2X, 3X) based on human-equivalent conversion. Exercise performance was assessed via grip strength and treadmill tests. Biochemical markers of fatigue, body composition, and safety indicators were also analyzed. Gut microbiota was evaluated using 16S rRNA sequencing and constrained principal coordinates analysis (CPCoA). Results: Four weeks of GU supplementation significantly enhanced exercise performance [(treadmill time ↑ Δ = 10.2-11.7 min versus vehicle (q ≤ 0.0002), grip strength ↑ Δ = 40.4-48.5 g (q ≤ 0.05)] and lean body mass [FFM ↑ at GU-1X (Δ = +0.80%, q = 0.0123)], surpassing the commercial BCAA control. Biochemical analyses indicated reduced exercise-induced lactate accumulation [(post-exercise lactate ↓ Δ = -2.71/-2.18 mmol·L^-1, q = 0.0006)]. Gut microbiota profiling revealed distinct shifts in community composition in GU-treated groups, notably with an increased abundance of beneficial taxa such as Lactobacillus and Muribaculum. These alterations reflect the prebiotic activity of seaweed-derived polysaccharides, promoting a healthier gut microbial profile. Notably, GU improved metabolic markers (aspartate aminotransferase, [AST]; lactate dehydrogenase, [LDH]) without inducing toxicity. Conclusions: These findings indicate that GU functions as a dual-action supplement, coupling amino acid-mediated muscle anabolism with microbiome modulation to enhance physical performance and metabolic health. As an upcycled marine product, it presents a sustainable and effective strategy for exercise support. Future studies should include 90-day safety, mechanistic assays, and a preregistered human pilot.

Keywords: Undaria pinnatifida; exercise performance; grouper bone; gut microbiota; microbiome modulation.

Figure 1

Time course of body weight changes during the 4-week intervention. Male ICR mice (n = 7 per group) were orally administered either vehicle, a commercial BCAA-positive control (PC), or the grouper bone/U. pinnatifida complex (GU-1X, GU-2X, or GU-3X) daily for 4 weeks. Body weight was measured at baseline and weekly thereafter, with a final measurement taken prior to euthanasia. Values are presented as mean ± SD.

Figure 2

Effect of GU supplementation on body composition. (A) Fat mass percentage and (B) free-fat mass percentage in male ICR mice after 4 weeks of treatment with vehicle (glucose water), PC (2050 mg/kg/day commercial BCAAs), GU-1X (2050 mg/kg/day GU), GU-2X (4100 mg/kg/day GU), or GU-3X (6150 mg/kg/day GU). Mice underwent an exercise performance test 1 h after the administered dose. Different letters (a, b, c) indicate statistically significant differences between groups (p < 0.05).

Figure 3

Effect of GU supplementation on exercise performance: forelimb grip strength (A) and treadmill exhausted time (B). Male ICR mice were pretreated for 4 weeks with vehicle (glucose water), PC (2050 mg/kg/day commercial BCAAs), GU-1X (2050 mg/kg/day GU), GU-2X (4100 mg/kg/day GU), or GU-3X (6150 mg/kg/day GU). Exercise performance tests were conducted 1 h after the administered dose. Different letters (a, b, c) indicate statistically significant differences between groups (p < 0.05).

Figure 4

Effect of GU supplementation on acute exercise-induced biochemical markers. (A) Serum lactate levels; (B) serum glucose levels. Male ICR mice were pretreated for 4 weeks with vehicle (glucose water), PC (2050 mg/kg/day commercial BCAAs), GU-1X (2050 mg/kg/day GU), GU-2X (4100 mg/kg/day GU), or GU-3X (6150mg/kg/day GU). Mice underwent exercise performance testing 1 h after the administered dose. Different letters (a, b) indicate statistically significant differences between groups (p < 0.05).

Figure 5

Effect of GU supplementation on tissue histology. Representative histological images of the liver (A), skeletal muscle (B), lung (C), kidney (D), heart (E), and EFP (F). Tissue sections were stained with hematoxylin and eosin (H&E) and observed under light microscopy at 200× magnification (A–F). EFP: epididymal fat pad.

Figure 6

The CPCoA plot visualizes the dissimilarities in microbial community composition across groups: vehicle, PC, GU-1X, GU-2X, and GU-3X. Each point represents an individual sample, with group-specific colors. Ellipses denote the 95% confidence interval for each group. The x-axis (CPCoA1) and y-axis (CPCoA2) explain 34.52% and 25.79% of the total variance, respectively. The spatial separation among groups indicates differences in gut microbial community structure in response to treatment.

Figure 7

Ternary plot illustrating the distribution of bacterial genera among three treatment groups.