Comparing the bacterial growth potential of ultra-low nutrient drinking water assessed by growth tests based on flow cytometric intact cell count versus adenosine triphosphate

Abstract

The bacterial growth potential (BGP) of drinking water is widely assessed either by flow cytometric intact cell count (BGP_ICC) or adenosine triphosphate (BGP_ATP) based methods. Combining BGP_ICC and BGP_ATP measurements has been previously applied for various types of drinking water having high to low growth potential. However, this has not been applied for water with ultra-low nutrient content, such as remineralised RO permeate. To conduct a sound comparison, conventionally treated drinking water was included in this study, which was also used as an inoculum source. BGP_ICC, BGP_ATP, intact cell-yield (Y_ICC), and ATP-yield (Y_ATP) were determined for conventionally treated drinking water (Tap-water) and remineralised RO permeate (RO-water). In addition, both BGP_ICC and BGP_ATP methods were used to identify the growth-limiting nutrient in each water type. The results showed that the BGP_ICC ratio between Tap-water/RO-water was ∼7.5, whereas the BGP_ATP ratio was only ∼4.5. Moreover, the Y_ICC ratio between Tap-water/RO-water was ∼2 (9.8 ± 0.6 × 10⁶ vs. 4.6 ± 0.8 × 10⁶ cells/µg-C), whereas the Y_ATP ratio was ∼1 (0.39 ± 0.12 vs. 0.42 ± 0.06 ng ATP/µg-C), resulting in a consistently higher ATP per cell in RO-water than that of Tap-water. Both BGP_ICC and BGP_ATP methods revealed that carbon was the growth-limiting nutrient in the two types of water. However, with the addition of extra carbon, phosphate limitation was detected only with the BGP_ICC method, whereas BGP_ATP was not affected, suggesting that a combination of carbon and phosphate is essential for biomass synthesis, whereas carbon is probably utilised for cellular activities other than cell synthesis when phosphate is limited. It was estimated that the intact cell-yield growing on phosphate would be 0.70 ± 0.05 × 10⁹ cells/µg PO₄-P.