Report on Test of XXXXX’s Immobilization of Daphnia

Portions redacted to protect confidentiality

Source:
XXXXXX 对蚤类运动抑制试验报告 1. 材料和方法1.1 供试样品信息（见表1）1.2 试验生物及来源大型水蚤， 从 XXXXXX 引种，本实验室条件下培养三代以上， 处于孤雌生殖状态。使用出生 6-24h 的幼蚤，健康、活泼。1.3 蚤类运动抑制试验1.3.1 试验时间：XXXX 年 X月X日— X  月X 日1.3.2 试验条件：试验用水经曝气去氯处理，pH值 6.0-9.0， 水中溶氧量为 4.0 mg/L 以上， 水质硬度 120-250 mg/L（以 CaCO3计），温度为 18-22 C， 光照 16L：8D。

1.3.3 试验设备：溶解氧测量仪、数字式 pH 计、温度计、 移液枪、容量瓶、玻璃棒、烧杯、滴管等。

1.3.4 染毒浓度及分组：

根据 XXXXXX 对大型水蚤运动抑制预试验结果，设置 12.0， 20.4， 34.6， 58.8， 100 mg a.i./L 5个浓度组，并设空白对照组，溶挤Tris-HCl 对照组和BSA蛋白（XXXXXXXXX）对照组， 处理组和对照组均设4 次重复，每个重复5 只试验用蚤。

 

1.3.5 染毒方法：

试验用 50mL 的烧杯加入 40mL 试验液，投放幼蚤5只。

1.3.6 症状观察与数据记录：

试验在人工气候箱中进行，观察试验用蚤的症状，记录处理后 24h 和 48h 受抑制数。

1.3.7 数据处理：

计算供试样品对大型水蚤 48h 的运动抑制率。

1.3.8 质量控制：

质量控制条件包括：

–对照组试验不活动蚤数少于10%。

–试验过程中供试物浓度不低于初始浓度的80%。

–在 20C 条件下， 参比物重铬酸钾对大型蚤EC50(24h）应处于 0.6-1.7 mg/L 之间。

–试验操作及试验过程中蚤不能离开水，转移时要用玻璃滴管。

 Target:
Report on Test of XXXXX’s Immobilization of Daphnia  1. Materials and Method1.1  Information on Test Sample (see Table 1)1.2  Test Organism and SourceDaphnia magna Straus, introduced from XXXXXXX. Under our laboratory conditions, we cultivated beyond third generation in parthenogenetic mode. We used young, healthy, lively Daphnia 6-24h old.1.3  Test of the Immobilization of Daphnia 1.3.1  Test period: XXXX – XXXX, XXXX1.3.2  Test conditions: The test used water that had been dechlorinated by aeration and had a pH of 6.0-9.0. The dissolved oxygen content of the water was above 4.0 mg/L. The water hardness was 120-250 mg/L (CaCO3 measurement). The temperature was 18-22 degrees C, the illumination 16L:8D.

1.3.3  Test equipment: Dissolved oxygen meter, digital pH meter, temperature gauge, pipettes, flasks, glass rods, beakers, and droppers.

1.3.4  Exposure concentration and grouping

Based on the results of pretests of the XXXXXX’s immobilization of Daphnia magna Straus, 5 concentration groups were set up: 12.0, 20.4, 34.6, 58.8, and 100 mg a.i./L. A blank control group, a Tris-HCl solvent control group, and a BSA (XXXXXXX) protein control group were also configured. Four iterations were conducted for all treatment and control groups, and each iteration used five test Daphnia.

1.3.5  Exposure method:

The test used 50mL beakers filled with 40mL of the test liquid, into which 5 young Daphnia were placed.

1.3.6   Symptom observation and data recording：

The test was conducted in an artificial climate box. Symptoms of the test Daphnids were observed and data on the numbers immobilized at 24h and 48h after treatment were recorded.

1.3.7  Data processing

The test sample’s rate of immobilization of Daphnia magna Straus over 48h was calculated.

1.3.8  Quality control

Quality control conditions included:

–Percentage of inactive Daphnia specimens in control group tests [should be] lower than 10%.

–Concentration of test substance [should be] no lower than 80% of that of the original concentration throughout the course of the test.

–At 20 degrees C, the EC50(24h) of the reference substance potassium dichromate on Daphnia magna Straus should be 0.6 – 1.7 mg/L.

—Daphnids should not be able to leave the water when the test is being conducted or throughout the course of the test, and glass droppers must be used when transferring Daphnia specimens.

 
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