In this study we applied bioassay using Chinese hamster ovary (CHO) cells with a heat shock protein (HSP) 47 promoter to the effluent of the wastewater treatment plants in Sapporo and we observed the statistically significant HSP production. This implied the effluent contained some organic matter which can stress the CHO cells. To investigate the possible causes of the toxicity of the effluent, we applied the assay to the rejected water from the sludge treatment plant, the mixtures of sewage and rejected water. The evolution of HSP production during the aerobic decay process and thickening process of sludge was also examined. These assay results showed that dissolved microbial products generated and/or released from activated sludge during its decay process in the aeration tank and during thickening and dewatering process in the sludge treatment train contributed to develop HSP production. The proteomics analysis was also applied to the effluent and detected the production of elongation factor 1β. This result implies that the effluent from wastewater treatment plants may cause changes in cell proteins involved in allergic reaction.