ISO 14669 pdf download.Water quality — Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea)
1 Scope
This International Standard describes a method for the determination of the acute toxicity to one of three specified species of marine copepod ( Copepoda, Crustacea ) of a) chemical substances which are soluble, or can be maintained as a stable suspension or dispersion, under the conditions of the test; b) industrial or sewage effluents, treated or untreated, after decantation, filtration or centrifugation if necessary; c) marine or estuarine waters.
2 Normative references
The following normative documents contain provisions which, through reference in this text, constitute provisions of this International Standard. For dated references, subsequent amendments to, or revisions of, any of these publications do not apply. However, parties to agreements based on this International Standard are encouraged to investigate the possibility of applying the most recent editions of the normative documents indicated below. For undated references, the latest edition of the normative document referred to applies. Members of ISO and IEC maintain registers of currently valid International Standards. ISO 5667-2, Water Quality — Sampling — Part 2: Guidance on sampling techniques . ISO 5667-1 6, Water Quality — Sampling — Part 1 6: Guidance on biotesting of samples.
3 Principle
Copepods are exposed to a range of concentrations in seawater of a chemical substance, effluent or water sample. Mortality of the copepods is recorded after 24 h and 48 h. The concentration which, in 48 h, kills 50% of exposed copepods under the conditions defined in this International Standard is determined. This concentration, known as the median lethal concentration, is designated 48 h LC50. NOTE If possible, the concentration which kills 50% of the exposed copepods in 24 h is also determined. This concentration is designated 24 h LC50. It may be appropriate for certain purposes to extend the exposure period to 96 h and to determine the 96 h LC50. If the method described in this International Standard is used for chemical substances, a limit test can be performed at 1 00 mg/l or at a lower concentration which is the maximum at which the substance is soluble or is in stable dispersion under the conditions of the test.
5 Reagents and materials
5.1 Test organism, one of the following species of marine copepod: a) Acartia tonsa Dana; b) Tisbe battagliai Volkmann-Rocco; c) Nitocra spinipes Boeck. Obtain the test organisms from laboratory cultures. Guidance on identification and culture methods for each species are given in Annex B. After hatching of eggs, the lifecycle of copepods consists of naupliar, copepodid and adult stages. The age and lifestage at the start of the test shall be indicated in the test report and are as follows: – Acartia tonsa : large copepodids (Stage 5) or adults; – Tisbe battagliai: copepodids 6 ± 2 days old; – Nitocra spinipes: adults 3 to 4 weeks old. 5.2 Dilution water. A natural or a synthetic seawater may be used as the dilution water. If natural seawater is used, it shall be collected from a location as distant as possible from known sources of pollution and filtered to remove indigenous organisms. If synthetic seawater is used, it shall be prepared by dissolving reagents of recognized analytical grade, or a commercially available formulation, in distilled or deionized water. However, for these copepod species, there is insufficient information on the use of synthetic seawater to allow a particular example to be recommended.The dilution water shall permit survival of the copepods for at least 48 h and should be from the same source as water that has been found to support culture of the organisms through at least two generations.