Deciphering the effects of temperature on bio-methane generation through anaerobic digestion

Anaerobic digestion (AD) is a sustainable wastewater treatment technology which facilitates energy, nutrient, and water recovery from organic wastes. The agricultural and industrial wastes are suitable substrates for the AD, as they contain a high level of biodegradable compounds. The aim of this study was to examine the AD of three different concentrations of phenol (100, 200, and 300 mg/L) containing wastewater with and without co-substrate (acetate) at four different temperatures (25, 35, 45, and 55 °C) to produce methane (CH₄)-enriched biogas. It was observed that the chemical oxygen demand (COD) and phenol removal efficiencies of up to 76% and 72%, respectively, were achieved. The CH₄ generation was found higher in anaerobic batch reactors (ABRs) using acetate as co-substrate, with the highest yield of 189.1 μL CH₄ from 500 μL sample injected, obtained using 200 mg/L of phenol at 35 °C. The results revealed that the performance of ABR in terms of degradation efficiency, COD removal, and biogas generation was highest at 35 °C followed by 55, 45, and 25 °C indicating 35 °C to be the optimum temperature for AD of phenolic wastewater with maximum energy recovery. Scanning electron microscopy (SEM) revealed that the morphology of the anaerobic sludge depends greatly on the temperature at which the system is maintained which in turn affects the performance and degradation of toxic contaminants like phenol. It was observed that the anaerobic sludge maintained at 35 °C showed uniform channels leading to higher permeability through enhanced mass transfer to achieve higher degradation rates. However, the denser sludge as in the case of 55 °C showed lesser permeability leading to limited transfer and thus reduced treatment. Quantitative real-time PCR (qPCR) analysis revealed a more noteworthy change in the population of the microbial communities due to temperature than the presence of phenol with the methanogens being the dominating species at 35 °C. The findings suggest that the planned operation of the ABR could be a promising choice for CH₄-enriched biogas and COD removal from phenolic wastewater.