Biological toxicity test method for evaluating the ecological safety of advanced oxidation processes

A method for evaluating ecological safety of an advanced oxidation process, comprising the following steps:

• (S1) Koi domestication:
• collecting tap water and dechlorinating the tap water by aeration for three days, selecting healthy Koi fish of 10±1 cm in length and keeping them in dechlorinated tap water for 10 days with a death rate lower than 3%, wherein the Koi fish is Cryprinus carpio haematopterus;
• (S2) exposure of the Koi fish to poison:
• (1) collecting waste water containing micro-contaminants or a secondary effluent from secondary sewage plants, dividing the collected waste water into two fractions, wherein the waste water in one fraction is subjected to an advanced oxidation process treatment and used as experimental waste water, wherein the waste water in another fraction is not subjected to the treatment and used as raw waste water, and wherein the advanced oxidation process treatment is UV/Na2S2O8, UV/H2O2, or UV/ClO2;
• (2) collecting tap water and dechlorinating the tap water as indicated in step S1, dividing dechlorinated tap water into two fractions, wherein the dechlorinated tap water in one fraction is subjected to the advanced oxidation process treatment and used as experimental tap water, and wherein the dechlorinated tap water in another fraction is not subjected to the treatment and used as raw tap water;
• (3) establishing three experimental groups and setting up one blank control group, wherein each group is provided with three parallel sub-groups, and each parallel sub-group is conducted with a fish tank, the size of which is unified to 50 L;
• wherein the first experimental group is a waste water control group with the experimental tap water added to each fish tank, the second experimental group is a waste water group with the raw waste water added to each fish tank, the third experimental group is a waste water experimental group with the experimental waste water added to each fish tank, and the blank control group is the tap water experimental group with the raw tap water added to each fish tank;
• wherein the three experimental groups and the blank control group have same conditions except for different water qualities;
• and wherein 1 μM non-steroid anti-inflammatory drug ibuprofen is added to each fish tank of the blank group, the first, second, and third experimental groups;
• and
• (4) putting 10 Koi fish, selected and domesticated with the raw tap water according to step S1, into each fish tank of the three experimental groups and the blank control group for exposure to poison for a period of 96-110 hours, wherein during the period of exposure to poison, half water in each fish tank is changed every day with water of same quality to maintain water quality in a stable condition;
• (S3) detection of activities of antioxidative enzymes:
• dissecting, after the exposure to poison according to step S2, the Koi fish in each group and extracting the liver, and then chopping and formulating the liver into a homogenate of liver tissues with a physiological saline according to a ratio of weight (g):
• volume (mL)=1:9;
• and determining the activities of antioxidative enzymes in the homogenate, wherein the antioxidative enzymes determined comprise superoxide dismutase, catalase, and glutathione peroxidase;
• and
• (S4) analysis of the enzyme activities of the Koi fish:
• statistically analyzing the enzyme activity results of step S3 to generate an average value of the enzyme activities for each of the three experimental groups and the blank control group, wherein the average value in each experimental or blank control group is compared to determine toxicity changes of waste water before and after the advanced oxidation process treatment;
• thereby evaluating the ecological safety of the advanced oxidation process.