Hormetic effect of dissolved organic matter from pig manure anaerobic digestion effluents on Chlorella sp.: Physiological and transcriptomic responses.

Microalgae-based biotreatment offers a sustainable and efficient solution for managing pig manure anaerobic digestion effluents (PMADE), a byproduct of intensive agricultural practices. Dissolved organic matter (DOM) in PMADE significantly influences microalgal physiological responses; however, the underlying mechanisms remain poorly understood. Here, we reveal a hormetic effect of DOM in PMADE on Chlorella sp. Moderate DOM concentrations (1000-1500 mg/L of COD) alleviated ammonium toxicity and promoted microalgal growth, whereas higher concentrations (>2000 mg/L of COD) exerted severe inhibitory effects. Through cascade extraction and targeted screening, we identified distinct impacts of five DOM fractions on Chlorella sp. Notably, low-molecular-weight acids and nitrogenous compounds, primarily dipeptides in the fractions, exhibited a promoting effect on growth by serving as carbon sources to enhance energy metabolism and protein synthesis. Among the fractions, antioxidant-active compounds further supported the growth by scavenging reactive oxygen species (ROS) and maintaining redox homeostasis. In contrast, fractions rich in alkanes and alkaloids, induced excessive ROS production under high concentrations, leading to impaired photosynthesis and severe DNA damage. Additionally, the substances such as humic acid and fulvic acid, as key components of PMADE, exhibited hormetic effects: at specific concentrations (humic acid<200 mg TOC/L; fulvic acid<100 mg TOC/L), they enhanced ROS scavenging, growth, photosynthesis and pigment accumulation, whereas exceeding these thresholds shifted their role from promotion to inhibition. Collectively, this study elucidates the dose-response relationship between DOM concentration and composition in relation to microalgal growth, identifies key DOM components influencing growth, and proposes targets for the selective removal of inhibitory DOM in PMADE.