Several single dose studies found that biochar was an effective additive promoting methanogenesis in anaerobic digestion. However, little attention was paid to the continuity of the biofilm formed on biochar and the subsequent evolution of microorganisms. In this study, the continuous performance and changes were quantified when biochar-biofilm consortia were applied as the recyclable inoculant to four cycles of acetate methanogenesis with increasing acetate loading. Biochar-biofilm consortia as the inoculant were found to consistently realize stable methane production, despite the removal of planktonic microorganisms in the reactor, and no extra inoculum and biochar were added. Consortia with biochar particles smaller than 5 μm promoted initiation of methanogenesis more rapidly than those with biochar particles larger than 1 mm, especially when the activity of microorganism was low. Moreover, the microorganisms were enriched throughout the continuous cycles. Biochar <5 μm was found to accumulate 6.6–7.1 E+11 16S copies per gram in the 3^rd and 4^th cycles, which was about 27−51 times that of biochar >1 mm and also comparable to a fresh sludge inoculum. In addition, specific functional methanogens proliferated during continuous application. Methanosarcina was dominant in biochar >1 mm, while the syntrophic acetate oxidizing bacteria Thermovirga and Mesotoga accounted for the majority of microorganisms in biochar <5 μm. Therefore, with the competitive total cell count dominated by functional microorganisms, biochar-biofilm consortia demonstrated feasible recycling and reuse for bioaugmentation purposes or in the economical long-term application of anaerobic digestion for waste or wastewater treatment.

几项单剂量研究发现，生物炭是促进厌氧消化中甲烷生成的有效添加剂。然而，很少关注在生物炭上形成的生物膜的连续性以及随后的微生物进化。在这项研究中，当将生物炭-生物膜联合体作为可循环接种物应用于醋酸盐甲烷生成的四个循环（醋酸盐负荷增加）时，其连续性能和变化得以量化。尽管已去除反应器中的浮游微生物，但发现作为接种剂的生物炭-生物膜联合会始终实现稳定的甲烷生产，并且未添加额外的接种物和生物炭。生物炭颗粒小于5微米的联盟比生物炭颗粒大于1毫米的联盟更快地促进了甲烷生成的开始，特别是当微生物的活性较低时。而且，微生物在整个连续循环中富集。发现3个以下的生物炭<5μm每克累积6.6–7.1 E + 11 16S拷贝^次和4^个循环，这是有关该生物炭的27-51倍> 1mm，并且还相当于一个新鲜污泥接种物。另外，在连续施用期间，特定的功能性产甲烷菌增殖。甲烷藻在> 1 mm的生物炭中占主导地位，而同食型乙酸盐氧化细菌Thermovirga和Mesotoga占<5μm的生物炭中的大多数微生物。因此，在功能性微生物主导的竞争性总细胞计数的情况下，生物炭-生物膜联盟证明了可行的回收利用和再利用，以用于生物强化目的或厌氧消化用于废物或废水处理的经济长期应用。