Henneberg Anja, Bender Katrin, Blaha Ludek, Giebner Sabrina, Kuch Bertram, Köhler Heinz-R, Maier Diana, Oehlmann Jörg, Richter Doreen, Scheurer Marco, Schulte-Oehlmann Ulrike, Sieratowicz Agnes, Ziebart Simone, Triebskorn Rita
Animal Physiological Ecology, University of Tübingen, Tübingen, Germany.
Department Aquatic Ecotoxicology, University of Frankfurt am Main, Frankfurt am Main, Germany.
PLoS One. 2014 Jun 5;9(6):e98307. doi: 10.1371/journal.pone.0098307. eCollection 2014.
Many studies about endocrine pollution in the aquatic environment reveal changes in the reproduction system of biota. We analysed endocrine activities in two rivers in Southern Germany using three approaches: (1) chemical analyses, (2) in vitro bioassays, and (3) in vivo investigations in fish and snails. Chemical analyses were based on gas chromatography coupled with mass spectrometry. For in vitro analyses of endocrine potentials in water, sediment, and waste water samples, we used the E-screen assay (human breast cancer cells MCF-7) and reporter gene assays (human cell line HeLa-9903 and MDA-kb2). In addition, we performed reproduction tests with the freshwater mudsnail Potamopyrgus antipodarum to analyse water and sediment samples. We exposed juvenile brown trout (Salmo trutta f. fario) to water downstream of a wastewater outfall (Schussen River) or to water from a reference site (Argen River) to investigate the vitellogenin production. Furthermore, two feral fish species, chub (Leuciscus cephalus) and spirlin (Alburnoides bipunctatus), were caught in both rivers to determine their gonadal maturity and the gonadosomatic index. Chemical analyses provided only little information about endocrine active substances, whereas the in vitro assays revealed endocrine potentials in most of the samples. In addition to endocrine potentials, we also observed toxic potentials (E-screen/reproduction test) in waste water samples, which could interfere with and camouflage endocrine effects. The results of our in vivo tests were mostly in line with the results of the in vitro assays and revealed a consistent reproduction-disrupting (reproduction tests) and an occasional endocrine action (vitellogenin levels) in both investigated rivers, with more pronounced effects for the Schussen river (e.g. a lower gonadosomatic index). We were able to show that biological in vitro assays for endocrine potentials in natural stream water reasonably reflect reproduction and endocrine disruption observed in snails and field-exposed fish, respectively.
许多关于水生环境中内分泌污染的研究揭示了生物群生殖系统的变化。我们采用三种方法分析了德国南部两条河流中的内分泌活动：（1）化学分析，（2）体外生物测定，以及（3）对鱼类和蜗牛的体内研究。化学分析基于气相色谱-质谱联用。对于水、沉积物和废水样品中内分泌潜力的体外分析，我们使用了E-筛检测法（人乳腺癌细胞MCF-7）和报告基因检测法（人细胞系HeLa-9903和MDA-kb2）。此外，我们用淡水螺蛳新西兰泥蜗对水和沉积物样品进行了生殖测试。我们将幼年褐鳟（Salmo trutta f. fario）暴露于废水排放口下游的水（舒森河）或参考站点的水（阿尔根河）中，以研究卵黄蛋白原的产生。此外，在两条河流中捕获了两种野生鱼类，圆腹雅罗鱼（Leuciscus cephalus）和点纹银鮈（Alburnoides bipunctatus），以确定它们的性腺成熟度和性腺指数。化学分析仅提供了关于内分泌活性物质的少量信息，而体外检测揭示了大多数样品中的内分泌潜力。除内分泌潜力外，我们还在废水样品中观察到了毒性潜力（E-筛检测法/生殖测试），这可能会干扰并掩盖内分泌效应。我们的体内测试结果大多与体外检测结果一致，揭示了两条被调查河流中一致的生殖干扰（生殖测试）和偶尔的内分泌作用（卵黄蛋白原水平），舒森河的影响更为明显（例如性腺指数较低）。我们能够证明，对天然溪水中内分泌潜力进行的生物体外检测分别合理地反映了在蜗牛和野外暴露鱼类中观察到的生殖和内分泌干扰情况。
