Estrogenic activity and estrogenic chemicals in landfill leachate were investigated by yeast two-hybrid assay and chemical analysis. Leachate sample extracted by liquid-liquid extraction with dichloromethane at pH 7.0 showed a higher dose-response curve than sample extracted at pH 3.0. or than sample extracted by solid phase extraction at either pH 7.0 or 3.0. The fraction extracted at pH 3.0 specifically inhibited not only growth of yeast but also estrogenic activity in this assay, suggesting that it contained anti-estrogenic chemicals. The greatest contributor to estrogenic activity among the chemicals identified in leachate extract was bisphenol A, with an estimated contribution ratio of 84%. The contribution ratios of 4-nonyl phenol (4-np) and 4-tert-octyl phenol (4-t-op) were estimated at 1.0%, and 0.1%, respectively, while natural estrogens such as 17beta-estradiol or estrone were below detection limit, so that their contribution ratio was estimated at no more than 10%. The estrogenic activity of leachate was decreased by aeration treatment alone after 7 days, and was no longer detected after 22 days. Concentrations of bisphenol A, 4-np and 4-t-op likewise decreased with aeration.
We previously developed a simplified yeast two-hybrid assay of estrogenic activity. In the present study, the optimal conditions for sensitivity and stability of this assay were decided. The assay could determine the estrogenic activity of 4 x 10(-11) mol dm(-1) 17beta-estradiol and also anti-estrogenic activity by using 4-hydroxytamoxifen as a control. The assay was used to test various chemicals suspected of estrogenicity. Many kinds of chemical, including alkylphenols, naphthols, xylenols, methoxychlor, phthalates, and bisphenol-A, showed estrogenic activity, and several, such as 2,5-xylenol and di-iso-octyl phthalate, showed anti-estrogenic but no estrogenic activity. Copresence of two estrogenic chemicals produced additive activity. The assay was also used to test leachate and water samples from the sea-based landfill of Osaka North Port in Japan. Estrogenic activity was detected in leachate from a municipal waste-dumping site, but not in that from a dredged soil-dumping site, which did however cause inhibition of cell growth.
In order to assess the degradability of plastics in solid waste disposal landfill sites, microbial populations capable of degrading five kinds of plastic-constituting polymers, poly epsilon-caprolactone (PCL), polylactic acid (PLA), polyethylene glycol (PEG), poly-beta-hydroxybutyrate (PHB) and cellulose acetate (CA), in a sea-based solid waste disposal site were investigated. Enumeration of aerobic and anaerobic polymers-degrading microorganisms (PDMs) was performed against to total 8 leachate samples, which were seasonally collected from the facultative pretreatment pond and the aerated lagoon. Both aerobic and anaerobic PDMs for natural polymers, PHB and CA, were found in all of the samples, while those for chemically-synthesized polymers, PCL, PLA and PEG, could not be always detected. In most cases, the ratios of the PHB- and CA-degraders to the heterotrophic bacterial population were more than 0.1%. On the other hand, the ratios of PCL-, PLA- and PEG-degraders were often much lower. These data indicate that the plastics degradation potential is commonly present in the studied disposal site, and that the degradation potential for plastics composed of chemically-synthesized polymers is inferior to that of natural polymers. Population sizes of the PDMs correlated to those of heterotrophic bacteria, and the counts of aerobic heterotrophic bacteria and PDMs in the aerated lagoon tended to be higher than those of anaerobic ones, indicating that the aeration of the leachate resulted in the activation of growth of whole aerobic microbial community including the PDMs.
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