Effect of Long-Term Freezing and Freeze–Thaw Cycles on Indigenous and Inoculated Microorganisms in Dewatered Blackwater

Abstract

Wastewater treatment in many Arctic regions is inadequate, even nonexisting. Natural freezing of wastewater in those areas may be beneficial for reduction of microorganisms. The aim of this study was to investigate the effect of long-term freezing, and repeated freezing and thawing, on indigenous coliforms, fecal streptococci, and antibiotic-resistant (AR) bacteria, and inoculated Salmonella Enteriditis and E. coli bacteriophage ΦX174 in dewatered blackwater. At the end of the long-term freezing experiment (10 months), an MPN recovery study was done, including the microbial groups that had shown the largest reduction, using tryptone soy broth at incubation temperatures of 10 and 20 °C overnight for the coliforms and AR bacteria, and buffered peptone water at incubation temperature of 37 °C for 18-20 h for Salmonella. Fecal streptococci were more resistant to long-term freezing than the coliform group. Total number of AR bacteria decreased slowly but constantly over the 10-month freezing period. Salmonella rapidly decreased and were nondetectable within a week but exhibited some recovery after 10 months of freezing, whereas limited or no recovery of coliforms and AR-bacteria was detected. Bacteriophages showed limited reduction during the long-term freezing. Repeated freezing and thawing increased the reduction of all tested microbial groups markedly.