The effect of limiting phosphorus (P) in activated sludge was investigated in laboratory-scale sequencing batch reactors (SBRs). Correlative microscopy revealed that P-limitation (COD:N:P=100:5:0.05) leads to morphological changes in floc structure and the composition of extracellular polymeric substances (EPS). This was found to be accompanied by expression of quorum-sensing in an acyl homoserine lactone bioassay. Differential gene expression in relation to P-limitation was examined in a global profile using the Affymetrix™ Escherichia coli antisense genomic microarray. Three separate experiments were conducted where the impact of P-limitation was examined under batch conditions and in SBRs at stable operating conditions and within 3–7 days following a down-shift in P. Significant changes in open reading frames (ORF) and intergenic regions based on the E. coli microarray were observed. Several genes associated with cell structure, including slt, wbbH, fimH, amB, rfaJ and slp were found to be expressed. Quorum regulated genes were also found to be expressed including psiF which is known to be induced by P-starvation (92% confidence level; 1.45 log ratio).

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