The use of next-generation diagnostic tools to optimise the anaerobic digestion of municipal sewage sludge has the potential to increase renewable natural gas recovery, improve the reuse of biosolid fertilisers and help operators expand circular economies globally. This review aims to provide perspectives on the role of microbial ecology in improving digester performance in wastewater treatment plants, highlighting that a systems biology approach is fundamental for monitoring mesophilic anaerobic sewage sludge in continuously stirred reactor tanks. We further highlight the potential applications arising from investigations into sludge ecology. The principal limitation for improvements in methane recoveries or in process stability of anaerobic digestion, especially after pre-treatment or during co-digestion, are ecological knowledge gaps related to the front-end metabolism (hydrolysis and fermentation). Operational problems such as stable biological foaming are a key problem, for which ecological markers are a suitable approach. However, no biomarkers exist yet to assist in monitoring and management of clade-specific foaming potentials along with other risks, such as pollutants and pathogens. Fundamental ecological principles apply to anaerobic digestion, which presents opportunities to predict and manipulate reactor functions. The path ahead for mapping ecological markers on process endpoints and risk factors of anaerobic digestion will involve numerical ecology, an expanding field that employs metrics derived from alpha, beta, phylogenetic, taxonomic, and functional diversity, as well as from phenotypes or life strategies derived from genetic potentials. In contrast to addressing operational issues (as noted above), which are effectively addressed by whole population or individual biomarkers, broad improvement and optimisation of function will require enhancement of hydrolysis and acidogenic processes. This will require a discovery-based approach, which will involve integrative research involving the proteome and metabolome. This will utilise, but overcome current limitations of DNA-centric approaches, and likely have broad application outside the specific field of anaerobic digestion.

The process of breaking down chicken manure through anaerobic digestion is an effective waste management technology. However, chicken manure can be a challenging feedstock, causing ammonia stress and digester instability. This study examined the impacts of adding wood biochar and acid-alkali-treated wood biochar to anaerobically digest chicken manure under conditions of ammonia inhibition. The results highlighted that only the addition of 5 % acid-alkali-treated wood biochar by volume can achieve cumulative methane production close to the typical methane potential range of chicken manure. The treated wood biochar also exhibited highest total ammonia nitrogen removal compared to the Control treatment. Scanning Electron Microscope revealed growing interactions between biochar and methanogens over time. Real-time polymerase chain reaction showed that treated wood biochar produced the highest number of bacterial biomass. In addition, 16S amplicon-based sequencing identified a more robust archaeal community from treated biochar addition. Overall, the acid-alkali treatment of biochar represents an effective method of modifying biochar to improve its performance in anaerobic digestion.

Anaerobic digestion has attracted great interest for use in the management of organic wastes and the production of biomethane. However, this process is facing challenges, such as a high concentration of ammonia nitrogen, which affects the methanogenesis process and, thus, the production of methane. This study investigates the use of biosolid-derived biochar for mitigating ammonia stress and improving methane production during the anaerobic digestion of chicken manure, using both pristine biochar and biochar modified with a potassium hydroxide (KOH) solution. Batch mesophilic anaerobic digestion (37 °C) was carried out over 18 days. When compared to chicken-manure-only controls, a significant increase in methane formation was observed in the digesters amended with biochar and KOH-modified biochar, producing 220 L kg⁻¹ volatile solids (VSs) and 262 L kg⁻¹ VSs of methane, respectively, compared to 139 L kg⁻¹ VSs from the control digesters. The use of biochar and KOH-modified biochar resulted in a significant reduction of 8 days in the lag phase. The total ammonia nitrogen (TAN) concentration was reduced in the digesters with biochar and KOH-modified biochar by 25% and 35.5%, respectively. The quantitative polymerase chain reaction (QPCR) data revealed that the number of 16S rRNA gene copies was around 50,000 and 41,000 times higher in the biochar and KOH-modified biochar digesters, respectively, compared to the control digesters on day 18. The taxonomic profiles indicated that the BC and KOH-BC digesters contained a mixture of methanogenic pathways, including acetoclastic (Methanosaetaceae), hydrogenotrophic (Methanosarcinaceae), and methylation (Methanofastidiosaceae). This mix of pathways suggests a more robust archaeal community and, hence, more efficient methanogenesis. The results show that the addition of biosolids biochar enhances anaerobic digestion, mitigates ammonia stress to methanogens, and significantly increases biogas production.