Effectiveness of layered inoculation in solid-state anaerobic co-digestion of pig manure and corn straw: Focus on macro-, micro-, and genetic-levels.

Layered inoculation can achieve rapid start-up and promote methanation performance of anaerobic digesters. Daily specific methane yield (SMY) rapidly increased to 2.93 mL/g VS/d during 0-13 days, and cumulative SMY reached 212 mL/g VS in the solid-state anaerobic co-digestion (SS-AcoD) of pig manure and corn straw. Data were collected at macro-, micro-, and genetic-levels of each substrate layer. The results showed that layered inoculation could improve volatile fatty acids utilization and prevent adverse effects of high total ammonium nitrogen concentrations. Layered inoculation accelerated hydrolysis, acidification, and methanogenesis of substrates, as evidenced by the efficient inoculation of Bacteroidetes, Anaerolineales, Methanosphaerula, and Methanothrix, which were primarily from inocula. The various stages of SS-AcoD were synergistically initiated during the first 13 days, and acetoclastic pathway was boosted. These results further explain why layered inoculation is an efficient method for improving methanation performance of SS-AcoD and achieving efficient utilization of organic solid waste.

Methanogenic community during the anaerobic digestion of different substrates and organic loading rates.

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-1  day-1 , 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.