ISO 14669-1999 PDF

Full title and description

ISO 14669:1999 — Water quality — Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea). This International Standard specifies a single‑species acute lethality bioassay using one of three named marine copepod species to determine 24 h/48 h (and optionally 96 h) LC50 values for chemical substances, industrial or sewage effluents and marine or estuarine waters.

Abstract

The standard describes procedures for exposing copepods to a series of concentrations in seawater and recording mortality after 24 h and 48 h to determine the 48 h median lethal concentration (48 h LC50). It covers use of chemical test substances (soluble or in stable dispersion), treated or untreated effluents (after decantation/filtration/centrifugation if necessary) and environmental water samples. Test design, controls (including solvent controls), preliminary and definitive tests, and calculation methods for LC50 (probit, moving average, binomial or graphical approaches) are specified.

General information

• Status: Published — International Standard; under review/to be revised (ISO review history shows confirmation in 2020 and stage updates indicating revision activity).
• Publication date: April 1999 (Edition 1, 1999-04).
• Publisher: International Organization for Standardization (ISO).
• ICS / categories: 13.060.70 — Examination of biological properties of water / water quality biotesting.
• Edition / version: Edition 1 (1999). Note: an ISO FDIS (final draft) for a revised edition (ISO/FDIS 14669) has appeared in the ISO lifecycle indicating a replacement in development.
• Number of pages: 16 pages (ISO catalogue listing).

Scope

Specifies a laboratory method to determine acute lethal toxicity to one of three specified marine copepod species (Acartia tonsa; Tisbe battagliai; Nitocra spinipes) for: (a) chemical substances that are soluble or can be kept in stable suspension/dispersion under test conditions; (b) industrial or sewage effluents (after appropriate pre‑treatment such as decantation, filtration or centrifugation where necessary); and (c) marine or estuarine waters. The standard sets minimum experimental conditions and acceptance criteria to generate 24 h and 48 h LC50 values (with optional 96 h values).

Key topics and requirements

• Design principle: preliminary range‑finding test followed by a definitive test to permit calculation of 48 h LC50 (24 h LC50 optional; 96 h LC50 may be appropriate in some cases).
• Test organisms: specified laboratory‑cultured copepod species — Acartia tonsa, Tisbe battagliai, Nitocra spinipes; required life stage and culture conditions are given.
• Environmental conditions: temperature control (recommended 20 °C ± 2 °C), 16 h/8 h light/dark photoperiod, specified salinity ranges and dissolved oxygen/pH requirements for dilution water.
• Replicates and organism density: guidance on minimum numbers (example: minimum 20 copepods per concentration, replicate containers recommended) and maximum densities to avoid crowding effects.
• Endpoints and calculations: mortality recorded at 24 h and 48 h (criteria for death specified); LC50 calculated using appropriate statistical methods (probit, moving average, binomial or graphical). Measurement and analytical checks on exposure concentrations are included with validity criteria (e.g., relative standard deviation limits).
• Controls and reference toxicant checks: use of controls, solvent controls if solvent is employed, and recommended reference toxicant (e.g., 3,5‑dichlorophenol) ranges to verify organism sensitivity.

Typical use and users

Used by environmental testing laboratories, ecotoxicologists, regulatory agencies, water quality laboratories and consultants for assessing acute toxicity of chemicals, effluents and environmental waters to marine invertebrates. Typical applications include environmental monitoring, effluent compliance testing, chemical hazard assessment and product safety evaluation where marine species sensitivity is relevant.

Related standards

Standards commonly referenced alongside ISO 14669 include ISO 5667‑16 (guidance on sampling and biotesting of samples) and other marine ecotoxicity methods such as ISO 10253 (marine algal growth inhibition test) and ISO 16712 (acute toxicity of marine sediment to amphipods). These documents provide complementary sampling, biotest selection and sediment/algal test procedures used in integrated water quality assessments.

Keywords

marine copepods, acute toxicity test, LC50, water quality, ecotoxicology, Acartia tonsa, Tisbe battagliai, Nitocra spinipes, bioassay, effluent testing, marine/estuarine waters.

FAQ

Q: What is this standard?

A: ISO 14669:1999 is an ISO International Standard that specifies a laboratory method for determining acute lethal toxicity to selected marine copepod species, producing 24 h/48 h (and optionally 96 h) LC50 results for chemicals, effluents and marine/estuarine waters.

Q: What does it cover?

A: It covers test organism selection and culture stage, test system and environmental controls (temperature, light cycle, salinity, oxygen and pH), test design (preliminary and definitive tests), replication and organism density, control and solvent control procedures, reference toxicant checks, endpoint definitions (mortality observation) and statistical methods for LC50 calculation.

Q: Who typically uses it?

A: Environmental laboratories, ecotoxicologists, regulatory agencies, consultants and industry testing groups conducting marine toxicity assessments or compliance testing that require standardized acute toxicity data for marine invertebrates.

Q: Is it current or superseded?

A: The 1999 edition is listed as published and was confirmed in ISO review cycles (last confirmation noted in 2020). ISO’s lifecycle indicates the standard is intended to be revised and an ISO/FDIS (final draft) for a revised edition (ISO/FDIS 14669) has been recorded as under development; users should check the ISO catalogue for the very latest publication status and any replaced/updated edition before relying on the method. (Dates and lifecycle notes from ISO catalogue: published April 1999; confirmation and review activity through 2020 and revision stage updates in 2023.)

Q: Is it part of a series?

A: It belongs to ISO activities on water quality and biological testing (ISO/TC 147/SC 5) and is used alongside other water‑quality biotesting and sampling standards (for example ISO 5667‑16 guidance on biotesting, ISO 10253 algal tests, ISO 16712 amphipod sediment tests).

Q: What are the key keywords?

A: Marine copepod, acute toxicity, LC50, bioassay, water quality, effluent testing, Acartia tonsa, Tisbe battagliai, Nitocra spinipes, ecotoxicology.