Objectives Dietary strategies to improve arachidonic acid:eicosapentaenoic acid (AA:EPA) ratios are of interest due to potential reductions in inflammation and oxidative stress following exercise. The aim of this study was to investigate the impact of a novel dietary intervention, that is, the ingestion of 30 g of dark chocolate, on blood lipid profiles and gut microbiota composition in elite male soccer players. Methods Professional male soccer players were randomly assigned to the experimental group (DC) provided with 30 g of dark chocolate or to the control group (WC), provided with 30 g of white chocolate, for 30 days. Before and after intervention, blood, fecal sample, and anthropometry data were collected. For each outcome, two-way repeated-measure analysis of variance was used to identify differences between baseline and endpoint (Week 4), considering treatment (dark chocolate, white chocolate) as intersubjects` factors. Metagenomic analysis was performed following the general guidelines, which relies on the bioBakery computational environment. Results DC group showed increased plasma polyphenols (from 154.7 ± 18.6 µg gallic acid equivalents/ml to 185.11 ± 57.6 µg gallic acid equivalents/ml, ? pre vs. post = +30.41 ± 21.50) and significant improvements in lipid profiles: total cholesterol (? -32.47 ± 17.18 mg/dl DC vs. ? -2.84 ± 6.25 mg/dl WC, Time × Treatment interaction p < .001), triglycerides (? -6.32 ± 4.96 mg/dl DC vs. D-0.42 ± 6.47 mg/dl WC, Time × Treatment interaction p < .001), low-density lipoprotein (D -18.42 ± 17.13 mg/dl vs. D-2.05 ± 5.19 mg/dl WC, Time × Treatment interaction p < .001), AA/EPA ratio (D -5.26 ± 2.35; -54.1% DC vs. D-0.47 ± 0.73, -6.41% WC, Time × Treatment interaction p < .001) compared with WC group. In addition, 4 weeks of intervention showed a significant increase in high-density lipoprotein concentration in DC group (D + 3.26 ± 4.49 mg/dl DC vs. ? -0.79 ± 5.12 mg/dl WC). Microbial communities in the DC group maintained a slightly higher microbial stability over time (exhibiting lower within-subject community dissimilarity). (D=Delta) Conclusion Ingesting 30 g of dark chocolate over 4 weeks positively improved AA:EPA ratio and maintained gut microbial stability. Dark chocolate ingestion represents an effective nutritional strategy to improve blood lipid profiles in professional soccer players.
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