Suitability of high-hardness COMBO medium for ecotoxicity testing using algae, daphnids, and fish
Suitability of high-hardness COMBO medium for ecotoxicity testing using algae, daphnids, and fish
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DOI：
10.1007/s001289900979
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发表时间：
1999-09-01
影响因子：
2.7
通讯作者：
Ling, Z
中科院分区：
文献类型：
--
作者：
Baer, KN;Ziegenfuss, MC;Ling, Z
材料和方法SD。 Magna Straus 培养物（从费城自然科学院获得的克隆 5）分别保存在含有 100 mL HH COMBO 培养基的 150 mL 玻璃烧杯中。使用分析级化学品和 Barnstead® Nanopure 超纯水制备培养基（表 1；Baer 和 Goulden，1998）。将培养基储存在 4°C 的黑暗中，并在使用前约 12 小时在环境室温下加热并剧烈通气。以75,000至100,000个细胞/ml的浓度每周3次向培养物喂食活绿藻A. falcatus。所有培养和测试均在20±2℃的恒温浴中进行，光照周期为16：8小时。黑头鲦鱼 (P. promelas) 幼鱼和胚胎/幼虫购自 Aquatic Biosystems, Inc., Fort Collins, CO，并在脱氯自来水中适应约 14 天或立即使用。藻类培养物在 HH COMBO 半连续培养物或 ASM 培养基中的通气静态培养物中生长。收获细胞并补充培养液以维持细胞处于对数生长期。氯化镉和氯化钾购自Fisher Scientific。所有测试溶液均在 HH COMBO 中制备并立即用于毒性测试。使用空气/乙炔火焰原子吸收分光光度计（Perkin Elmer，型号 3030B）在选定的时间段测量镉。该方法的检测限约为 0.050 mg/L。测试溶液中高于检测限的镉回收率为 85% 至 106%。
MATERIALS AND METHODSD. magna Straus cultures (clone 5 obtained from Academy of Natural Sciences of Philadelphia) were individually held in 150 mL glass beakers containing 100 mL, of HH COMBO medium. The medium was prepared using analytical grade chemicals and Barnstead® Nanopure ultrapure water (Table 1; Baer and Goulden 1998). Medium was stored in the dark at 4 C and warmed in ambient room temperature with vigorous aeration approximately 12 hr prior to use. Cultures were fed live green algae, A. falcatus, at a concentration of 75,000 to 100,000 cells/ml three times weekly. All culturing and testing were performed in a constant temperature bath at 20±2 C with a 16: 8-hr light: dark photoperiod. Fathead minnows (P. promelas) juvenile and embryos/larvae were purchased from Aquatic Biosystems, Inc., Fort Collins, CO and acclimated in dechlorinated tap water for~ 14 days or used immediately. Algal cultures were grown in semicontinuous cultures of HH COMBO or aerated static cultures in ASM medium. Cells were harvested and culture solution replenished to maintain cells in log growth phase.Cadmium chloride and potassium chloride were purchased from Fisher Scientific. All test solutions were prepared in HH COMBO and used immediately in toxicity tests. Cadmium was measured at selected time periods using an air/acetylene-flame atomic absorption Spectrophotometer (Perkin Elmer, Model 3030B). The detection limit for this method is approximately 0.050 mg/L. Recoveries of cadmium in test solutions above the detection limit ranged from 85 to 106%.