Evaluation of the inactivation of human Coxsackievirus by thermophilic and mesophilic anaerobic digestion using integrated cell culture and reverse transcription real-time quantitative PCR

• Inactivation of human Coxsackievirus using thermophilic and mesophilic anaerobic digestion.
• Integrated cell culture and real-time PCR for assessing disinfection of the virus.
• 7 log reduction of viral genome in 2 days of TAD treatment while 2.6 log in 14 days of MAD treatment.
• A synergistic effect of microbes, proteinase, RNase, and temperature for viral inactivation.
• TAD process is a biological way to achieve pathogen control in wastewater and biowaste.

The virucidal effects of anaerobic digestion were evaluated using human Coxsackievirus as a model for the Enterovirus family. Coxsackievirus was inactivated completely by thermophilic anaerobic digestion (TAD). By 4 h no living and infectious virus remained and no detectable viral RNA was present after 2 days in TAD (7.0 log reduction). Compared to TAD, 2.6 log reduction of viral RNA was achieved by 14 days in mesophilic anaerobic digestion (MAD) (p < 0.0001). Although cytopathogenic effect was not observed in the cultured cells, low levels of intracellular viral RNA were detected after one day of MAD treatment indicating that Coxsackievirus had infected the cells but could not replicate. The combination of thermal and biochemical effects in TAD plays a critical role for viral disinfection. The results of this study indicate that selection of the right configuration of anaerobic digestion for treatment of biowaste may reduce the risk of viral contamination to the environment and water source.

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