【關(guān)鍵詞】Anaerobic digestion ,Salinity stress ,Conductive materials ,Direct interspecies electron transfer ,Metabolic reconstruction
【DOI】https://doi.org/10.1016/j.envres.2023.115779
【影響因子(IF)】8.43
【出版期刊】《Environmental Research》
【產(chǎn)品原文引用】
Flow cytometry (BD, AccuriTMC6 plus, USA) was employed with Annexin V-FITC/PI as the fluorescence dye to detect the intact and injured percentage of microbial cells (Gao et al., 2022). Take 10 mL sludge sample into a centrifuge tube, sonicate the sludge sample to produce a single-cell mixture, filter the single-cell mixture with a 40 μm cell filter to remove large particles, wash twice with phosphate buffered solution (PBS), and adjust the cell concentration to 1.0 × 106–1.0 × 108 cells/mL with PBS to make a cell test solution. Then 490 μL of cell test solution was added to a 2 mL centrifuge tube, 5 μL of Annexin V-FITC and 5 μL of PI dye were added in sequence and mixed evenly. Incubate for 15 min at room temperature in the dark, and the cultured samples were tested by flow cytometry. The data of the flow cytometer was analyzed by FlowJo software. Protease, ATP, acetate kinase (AK), and Coenzyme F420 activity were determined by ELISA kits (Hengyuan biological, Shanghai, China). Electron transport system activity (ETS) of AD sludge was found to be an effective indicator of overall microbial respiration activity in the AD process. The ETS was quantified using a modified 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride (INT) method and previously described (Tian et al., 2017). The microbial morphology was characterised by scanning electron microscopy (ZEISS Gemini SEM 300, Germany).
