A schematic drawing of the integrated process is shown in Fig. 1. The FWHB was mixed
with primary wastewater with a ratio of 1:4 (v/v) and used as the culture media for R.
glutinis and C. curvatus. Five milliliters of sub-cultured cells was used to inoculate into
50 mL culture medium in a 250-mL Erlenmeyer flask. The flask was incubated at 25 °C in
an orbital shaker set to170 rpm for 6 days. The initial pH in the medium was 6.0, and it was
very stable during the culture process. After this incubation, 90% of the cell suspension was
harvested and the produced biomass was separated by centrifugation. The supernatant was
inoculated with the 10% remaining cell suspension and further grown with nutrients under
the same culture conditions. The effluent from this second step was used to support growth
of the phototrophic algae Chlorella sorokiniana (UTEX 1602), which was possible since
the effluent still contained nitrogen and phosphorous. The seed cells of C. sorokiniana were
maintained in phototrophic culture in Kuhl’s medium [16], and the inoculation rate of the
yeast culture to the effluent was 1% (v/v). The pH of Kuhl’s medium was 6.0, and no pH
adjustment is necessary before and after the inoculating