| Summary: | In Taiwan, the mushroom <i>Antrodia camphorata</i> (AC) is used for medicinal purposes, including cancer prevention and treatment. This study aimed to investigate the effect of AC-supplemented diets on the innate immunity and disease resistance of the orange-spotted grouper <i>Epinephelus coioides</i> against two bacterial infections—<i>Vibrio alginolyticus</i> and <i>Streptococcus iniae.</i> Orange-spotted groupers were fed AC-supplemented diets at 0 (control), 1.5, 3.0, 4.5, and 6.0 g kg<sup>−1</sup> for 28 days and examined for innate immune responses and survival rates against <i>V. alginolyticus</i> and <i>S. iniae</i>. The results showed that the innate cellular and humoral immune parameters, including respiratory burst, phagocytic activity, and lysozyme activity of leukocytes, along with serum alternative complement activity (ACH<sub>50</sub>) and superoxide dismutase activity, as well as interleukin (IL)-4 and IL-6, were all significantly increased in groupers fed AC-supplemented diets. Groupers fed diets containing AC at doses higher than 3.0 g kg<sup>−1</sup> had better survival rates than the control group 3–18 days after <i>V. alginolyticus</i> or <i>S. iniae</i> challenge. The enhanced disease resistance may be attributable to AC-induced innate immunity, suggesting that utilizing AC as a dietary supplement at 3.0 g kg<sup>−1</sup> may enhance the disease resistance of orange-spotted groupers against Vibrio or Streptococcus infection.
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