Impact of calcium carbonate and temperature on survival of Escherichia coli in soil

Abstract

Spreading of waste organic matter on agricultural lands is considered to enhance soil microbial activities and physical properties and improves soil nutrient status. However, organic wastes have also been shown to be a source of microbial contaminants including pathogens. Related risks are governed by pathogens' survival and transport particularities. We evaluated the significance of high levels of CaCO3, common in arid and semi-arid soils, on survival of Escherichia coli NAR at different temperatures. Amendments of 0, 5, 10, 15 or 25 g CaCO3 were mixed into variable soil amounts to obtain 100 g soil-CaCO3 mixtures. Both sterile and non-sterile soil mixtures were tested. Suspensions of a nalidixic acid-resistant E. coli strain (E. coli NAR) were added to the mixtures at a rate of 10(6) cell g(-1) soil. Mixtures were incubated at 4, 15, or 37 °C at the soil's field capacity for water (i.e. 0.13 g g(-1)). Each treatment was tested in triplicate. Persistence of culturable E. coli NAR was verified throughout the incubation period. The recovery rates of culturable E. coli NAR were significantly correlated to CaCO3 concentrations (P < 0.05). Incubation temperature (T) was the most significant factor (P < 0.01). In non-sterile mixtures the largest decline in survival rates of E. coli NAR was measured for treatments with larger CaCO3 content (i.e. 15 and 25%). Interaction of temperature and CaCO3 was significant for E. coli NAR die-off. Sterilization of soil caused non-uniform fluctuations in the effect of treatments. The maximum calculated decay rate for E. coli NAR was 0.83 d(-1) for the 15 g CaCO3 non-sterile mixture incubated at 37 °C while the minimum was 0.09 d(-1) for the control unamended sterile soil incubated at 15 °C. A combination of high temperature, large CaCO3 concentrations and a non-sterile, biologically active soil created the least favorable conditions for E. coli survival.