Reduced EBPR performance in full and bench-scale EBPR studies was linked to the proliferation of GAOs but often time with the lack of any evidence. In this study, a detailed enzymatic study was coupled with batch tests and electron microscopy results for a realistic explanation. The results eliminated the possibility of population shift from PAO to GAO or other non-PAO due to the short batch test period provided which would not allow a population shift and further justified with the electron microscopy results. The results indicate that glycogen serves not only as source of reducing power for PHA production but also serves as an alternative energy source when the poly-P pool of the PAOs is depleted. Slow generation of ATP via glycolytic pathway at 5°C cannot satisfy energy requirements of EBPR cells to complete several cell functions including acetate uptake and PHA storage. However, the glycolytic pathway is efficiently operable at warm temperatures (>20°C). The reduced performance of enhanced EBPR facilities operated at warm temperature may not be a result of GAO proliferation; instead it may be related the efficient use of the glycolytic pathway by PAOs which results in more glycogen storage and less P uptake, thereby reducing the EBPR performance.

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