Types of Tests
Acute tests are short-term exposure tests (hours or days) and generally use lethality as an endpoint. In acute exposures, organisms come into contact with higher doses of the toxicant in a single event or in multiple events over a short period of time and usually produce immediate effects, depending on absorption time of the toxicant. These tests are generally conducted on organisms during a specific time period of the organism’s life cycle, and are considered partial life cycle tests. Acute tests are not valid if mortality in the control sample is greater than 10%. Results are reported in EC50, or concentration that will effect fifty percent of the sample size.
Chronic tests are long-term tests (weeks, months years), relative to the test organism’s life span (>10% of life span), and generally use sub-lethal endpoints. In chronic exposures, organisms come into contact with low, continuous doses of a toxicant. Chronic exposures may induce effects to acute exposure, but can also result in effects that develop slowly. Chronic tests are generally considered full life cycle tests and cover an entire generation time or reproductive life cycle (“egg to egg”). Chronic tests are not considered valid if mortality in the control sample is greater than 20%. These results are generally reported in NOECs (No observed effects level) and LOECs (Lowest observed effects level).
Early life stage tests are considered as subchronic exposures that are less than a complete reproductive life cycle and include exposure during early, sensitive life stages of an organism. These exposures are also called critical life stage, ebryo-larval, or egg-fry tests. Early life stage tests are not considered valid if mortality in the control sample is greater than 30%.
Short-term sublethal tests are used to evaluate the toxicity of effluents to aquatic organisms. These methods are developed by the EPA, and only focus on the most sensitive life stages. Endpoints for these test include changes in growth, reproduction and survival. NOECs, LOECs and EC50s are reported in these tests.
Bioaccumulation tests are toxicity tests that can be used for hydrophobic chemicals that may accumulated in the fatty tissue of aquatic organisms. Toxicants with low solubilities in water generally can be stored in the fatty tissue due to the high lipid content in this tissue. The storage of these toxicants within the organism may lead to cumulative toxicity. Bioaccumulation tests use bioconcentration factors (BCF) to predict concentrations of hydrophobic contaminants in organisms. The BCF is the ratio of the average concentration of test chemical accumulated in the tissue of the test organism (under steady state conditions) to the average measured concentration in the water.
Freshwater tests and saltwater tests have different standard methods, especially as set by the regulatory agencies. However, these tests generally include a control (negative and/or positive), a geometric dilution series or other appropriate logarithmic dilution series, test chambers and equal numbers of replicates, and a test organism. Exact exposure time and test duration will depend on type of test (acute vs. chronic) and organism type. Temperature, water quality parameters and light will depend on regulator requirements and organism type.
Effluent toxicity tests are tests conducted under the Clean Water Act, National Pollutant Discharge Elimination System (NPDES) permit program and are used by dischargers of contaminated effluent to monitor the quality of effluent into receiving waters. Acute Effluent Toxicity Tests are used to monitor the quality of industrial effluent monthly using acute toxicity tests. Effluent is used to perform static-acute multi concentration toxicity tests with C. dubia and P. promelas. The test organisms are exposed for 48 hours under static conditions with five concentrations of the effluent. Short-term Chronic Effluent Toxicity Tests are used to monitor the quality of municipal wastewater treatment plants effluent quarterly using short-term chronic toxicity tests. The goal of this test is to ensure that the wastewater is not chronically toxic. The major deviation in the short-term chronic effluent toxicity tests and the acute effluent toxicity tests is that the short-term chronic test lasts for seven days and the acute test lasts for 48 hours.
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