title: Development and assays estradiol equivalent concentration from prawn (p-EEQ) in river prawn, Macrobrachium nipponense, in Taiwan.
abstract: The present study established a fast and convenient bioassay method for aqueous ecosystems using the prawn estradiol equivalent concentration (p-EEQ) of male Macrobrachium nipponense, which produce vitellogenin (VTG) after exposure to xeno-estrogens. This method was then used to determine the concentrations of xeno-estrogen pollutants in the rivers of Taiwan. To establish the calibration curve for the concentrations based on the p-EEQ, the induced VTG content was determined using the alkali-labile phosphate method after male M. nipponense were exposed to 0, 10, 100, 1,000 and 10,000ng/L of 17β-estradiol for 1, 3, 5, 7, 10 and 14 days, respectively. The results of the experiments showed that the induced VTG content in all of the experimental groups stabilized after 10 days, except for the 10,000ng/L experimental group, in which the induced VTG content decreased after 10 days. A 17β-estradiol-VTG10dayresponse curve was then established based on the induced VTG content in the 0, 10, 100 and 1000ng/L experimental groups at day 10. After establishing the curve, male M. nipponense were captured from the upper, middle and lower reaches of the Chuo-shui River, the Beigang River, the Jishui River, the Agongdian River and the Sichong River in Taiwan, and the VTG content in these prawns was determined. In addition, the p-EEQ in the waters was determined based on the VTG content, and the estradiol equivalent concentration (EEQ) in the waters was also measured immediately after sampling using the solid-phase extraction-enzyme-linked immunosorbent assay (SPE-ELISA) method. The results showed that the p-EEQ in the middle and lower reaches of the rivers in certain parts of Taiwan ranged from 38 to 400ng/L, and the detection rate was 100%. Moreover, the EEQ ranged from 7.9 to 92.9ng/L, and the detection rate was 42.9%, indicating that most of the middle and lower reaches of the rivers in Taiwan were polluted by xeno-estrogens. The 17β-estradiol concentrations determined based on the p-EEQ were all higher than those based on the EEQ (SPE-ELISA method). The results of the present study showed that the use of M. nipponense to determine the p-EEQ in environmental waters provided advantages that included a high detection rate, high sensitivity and convenience. However, the p-EEQ cannot be used in waters that do not contain M. nipponense.
authors.affiliation: National Museum of Marine Biology & Aquarium, Pingtung, Taiwan, China; Graduate Institute of Marine Biology, National Dong Hwa University, 2 Houwan Road, Checheng, Pingtung 944, Taiwan, China.
authors.forename: Yuh-Wen
authors.initials: YW
authors.lastname: Chiu
authors.affiliation: Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung 80701, Taiwan, China.
authors.forename: Fang-Ling
authors.initials: FL
authors.lastname: Yeh
authors.affiliation: Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung 80701, Taiwan, China.
authors.forename: Bao-Sen
authors.initials: BS
authors.lastname: Shieh
authors.affiliation: Department of Environmental Resources Management, Chia Nan University of Pharmacy & Science, Tainan, Taiwan, China.
authors.forename: Chien-Min
authors.initials: CM
authors.lastname: Chen
authors.affiliation: Department of Aquatic Biosciences, National Chiayi University, 300 University Rd., Chiayi 60004, Taiwan, China.
authors.forename: Hong-Thih
authors.initials: HT
authors.lastname: Lai
authors.affiliation: Graduate Institute of Biotechnology, Chinese Culture University, Yang Ming Shan, Taipei, Taiwan, China.
authors.forename: Shu-Yin
authors.initials: SY
authors.lastname: Wang
authors.affiliation: Department of Environmental Resources Management, Chia Nan University of Pharmacy & Science, Tainan, Taiwan, China. Electronic address: daji@ms19.hinet.net.
authors.forename: Da-Ji
authors.initials: DJ
authors.lastname: Huang
mesh_terms: D000818:Animals; D001681:Biological Assay; D004305:Dose-Response Relationship, Drug; D004784:Environmental Monitoring / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D004958:Estradiol / Q000633:toxicity*; D004967:Estrogens / Q000633:toxicity*; D008297:Male; D033541:Palaemonidae / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D045483:Rivers / Q000737:chemistry*; D013624:Taiwan; D014819:Vitellogenins / Q000378:metabolism; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D004967:Estrogens; D014819:Vitellogenins; D014874:Water Pollutants, Chemical; D004958:Estradiol
keywords: Estradiol equivalent concentration form prawn; Macrobrachium nipponense; Vitellogenin