Ensuring the biological stability of drinking water is essential for modern drinking water supply. To understand and manage the biological stability, it is critical that the bacterial growth in drinking water can be measured. Nowadays, advance treatment technologies, such as reverse osmosis (RO), are increasingly applied in drinking water purification where the produced water is characterized by low levels of nutrients and cell counts. The challenge is, therefore, how to measure the low bacterial growth potential (BGP) of such ultra-pure water using the available methods which were originally developed for conventionally treated drinking water. In this study, we proposed a protocol to assess BGP of ultra-pure drinking water produced by RO and post-treatment (including remineralization). Natural bacterial consortium from conventional drinking water was added to all water samples during this study to ensure the presence of a wide range of bacterial strains. The method development included developing an ultra-pure blank with high reproducibility to lower the detection limit of the BGP method (50 ± 20 × 10 intact cells/mL) compared with conventional blanks such as bottled spring water, deep groundwater treated by aeration and slow sand filtrate of surface water supply. The ultra-low blank consists of RO permeate after adjusting its pH and essential mineral content under controlled laboratory conditions to ensure carbon limitation. Regarding the test protocol, inoculum concentrations of >10 × 10 intact cells/mL may have a significant contribution to the measured low levels of BGP. Pasteurization of water samples before measuring BGP is necessary to ensure reliable bacterial growth curves. The optimized method was used to assess BGP of ultra-pure drinking water produced by RO membranes and post-treatment (including remineralization), where the BGP has decreased more than 6-fold to a level of 90 ± 20 × 10 intact cells/mL compared with conventionally treated water (630 ± 70 × 10 intact cells/mL).
Measuring bacterial growth potential (BGP) involves sample pre-treatment and inoculation, both of which may introduce contaminants in ultra-low nutrient water (e.g., remineralized RO permeate). Pasteurization pre-treatment may lead to denaturing of nutrients, and membrane filtration may leach/remove nutrients into/from water samples. Inoculating remineralized RO permeate samples with natural bacteria from conventional drinking water leads to undesired nutrient addition, which could be avoided by using the remineralized RO permeate itself as inoculum. Therefore, this study examined the effect of pasteurization and membrane filtration on the BGP of remineralized RO permeate. In addition, the possibility of using bacteria from remineralized RO permeate as inoculum was investigated by evaluating their ability to utilize organic carbon that is readily available (acetate, glucose) or complex (laminarin, gelatin, and natural dissolved organic carbon), as compared with bacteria from conventional drinking water. The results showed that membrane filtration pre-treatment increased (140-320%) the BGP of remineralized RO permeate despite the extensive soaking and flushing of filters (>350 h), whereas no effect was observed on the BGP of conventional drinking water owing to its high nutrient content. Pasteurization pre-treatment had insignificant effects on the BGP of both water types. Remineralized RO permeate bacteria showed limitations in utilizing complex organic carbon compared with bacteria from conventional drinking water. In conclusion, the BGP bioassay for ultra-low nutrient water (e.g., remineralized RO permeate) should consider pasteurization pre-treatment. However, an inoculum comprising bacteria from remineralized RO permeate is not recommended as the bacterial consortium was shown to be limited in terms of the compounds they could utilize for growth.
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