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Appl Environ Microbiol. 1989 February; 55(2): 401-405

Simple method for the detoxification of wastewater ultrafiltration concentrates for rotavirus assay by indirect immunofluorescence.

J I Oragui and D D Mara

Department of Civil Engineering, University of Leeds, United Kingdom.

ABSTRACT

A simple method for the detoxification of ultrafiltration concentrates of wastewaters for rotavirus assay by the indirect immunofluorescence technique has been developed. Polyacrylamide (Bio-Gel) or dextran (Sephadex G50) beads were mixed with concentrates (0.5 g/10 ml, wt/vol) of wastewaters seeded with simian rotavirus SA11 and allowed to stand for 2 h. The supernatant was decontaminated with antibiotics and then assayed for rotaviruses. Concentrates from raw sewage and treated effluents seeded with SA11 were used to infect MA104 or LLC MK2 cell lines. The concentrates, particularly those from raw sewage and anaerobic waste stabilization ponds, were very toxic to the tissue culture cells. These toxic effects were determined by the detachment and subsequent loss of cells after incubation with concentrates and assay medium for 24 h. They were either completely eliminated or were reduced by greater than 80% after treatment with beads.

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Appl Environ Microbiol. 1989 February; 55(2): 401-405