Immunological methods for the study of zoogloea strains in natural environments

Since Zoogloea ramigera has been considered to be important in aerobic wastewater treatment, we have evaluated several methods for detecting and enumerating Z. ramigera in water and wastewater samples. Indirect immunoassay methods for the detection of Zoogloea strains were developed using polyclonal antibodies against the cells or the isolated exocellular polymer (EP) of the neotype Zoogloea ramigera strain 106 (ATCC 19544). The primary antibodies reacted with the cells and the exopolymer associated with finger-like zoogloeal projections, but not with other bacteria from natural samples. These antibodies allowed detection of Z. ramigera in environmental samples. Scanning electron microscopy (SEM) was used to show that the cells and the exocellular polymer of naturally occurring zoogloeal projections are antigenically and structurally related to those of Z. ramigera 106. Both immunological procedures and probes complementary to regions on the 16S rRNA could detect Z. ramigera in natural samples but the immunological procedures were easier to use. RT-PCR was also used to detect Z. ramigera in natural samples. These methods were also used to identify Z. ramigera in biofilms that developed over wastewater samples as part of an MPN procedure that was used to quantitate Z. ramigera at different stages of the wastewater treatment process and in different lakes. Z. ramigera could be found in all stages of wastewater treatment processes, from raw wastewater to chlorinated effluent. The highest concentration of Z. ramigera was found in the mixed liquor stage of the a wastewater treatment plant. Additionally, Z. ramigera was found in all eutrophic and mesotrophic lakes and in some oligotrophic lakes.