Three anaerobic reactors using pig manure (PM), maize straw (MS), and a mixture of the two as substrates were compared for archaeal community structure and diversity, and for methanogens response to increased organic loading rate (OLR, expressed in the mass of volatile solid (VS)). Methanogenic archaeal richness during codigestion of pig manure with maize straw (ACE: 2412) was greater than that during the others (ACE: 1225, 1467) at an OLR of 4 g L day , accompanied by high specific methane yield. Euryarchaeota and Crenarchaeota predominated during overall digestion of different substrates; with relative abundances of 63.5%-99.0% and 1.0%-36.3%, respectively. Methanosarcina was the predominant genus that accounted for 33.7%-79.8% of the archaeal community. The diversity in the PM digester decreased with increase in OLR, but increased in the MS digester. The diversity was stable during the codigestion with increased OLR. The relative abundances of hydrogenotrophic methanogens increased by 2.6 and 2.1 folds; the methanogenic community shifted from acetoclastic to hydrogenotrophic methanogens during digestion of MS, and of the mixture of MS and PM. Canonical correspondence analysis revealed a strong relationship between reactor parameters and methanogenic community.
Solid-state anaerobic digestion is easily inhibited by high volatile fatty acid induced by high total solids, although it is a promising technology. Previous studies on volatile fatty acid inhibition mainly focused on total solid content, co-digestion substrates, and external additives. The present study proposed a new inoculation method named layer inoculation and compared it to premixing inoculation in the solid-state anaerobic digestion of pig manure and maize straw. The results showed that the cumulative CH4 yields from layer inoculation (211.5 mL/g-VS) were 5.64 times more than premixing inoculation (37.5 mL/g-VS) under a low inoculation ratio (25%), with the values of total volatile fatty acid being greater than 30.0 mg/g. The concentrations of total VFAs and acetic acid from layer inoculation decreased dramatically during days 18–43. Layer inoculation also showed wider specific methane yield peaks and shorter startup times than premixing inoculation. Methanosphaerula and Methanothrix were the most dominant genera, while the genus Methanosphaerula did not correlate with volatile fatty acids, pH, or total ammonia nitrogen. The hydrogenotrophic methanogen pathway was predominant during solid-state anaerobic digestion; the shift from hydrogenotrophic to acetoclastic occurred in premixing inoculation, and it was stable in layer inoculation (61.20–68.88%). Overall, layer inoculation can effectively enhance methane production under high volatile fatty acid concentrations compared with premixing inoculation.
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