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.
The spread of epidemically successful nontyphoidal Salmonella clones has been suggested as the most important cause of salmonellosis in industrialized countries. Factors leading to the emergence of success clones are largely unknown, but their ability to survive and grow after physical stress may contribute. During epidemiological studies, a mathematical model was developed that allowed estimation of a factor (q) accounting for the relative ability of Salmonella serovars with different antimicrobial resistances to survive in the food chain and cause human disease. Based on this q-factor, 26 Salmonella isolates were characterized as successful or nonsuccessful. We studied the survival and growth of stationary- and exponential-phase cells of these isolates after freezing for up to 336 days in minced meat. We also investigated survival and growth after dehydration at 10°C and 82% relative humidity (RH) and 25°C and 49% RH for 112 days. Stationary-phase cells were reduced by less than 1 log unit during 1 year of freezing, and growth was initiated with an average lag phase of 1.7 h. Survival was lower in exponential-phase cells, but lag phases tended to be shorter. High humidity and low temperature were less harmful to Salmonella than were low humidity and high temperature. Tolerance to adverse conditions was highest for Salmonella Infantis and one Salmonella Typhimurium U292 isolate and lowest for Salmonella Derby and one Salmonella Typhimurium DT170 isolate. Dehydration, in contrast to freezing, was differently tolerated by the Salmonella strains in this study, but tolerance to freezing and dehydration does not appear to contribute to the emergence of successful Salmonella clones.
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