SVENSK STANDARD
Fastställd/Approved: 2016-12-08 Publicerad/Published: 2016-12-22 Utgåva/Edition: 3
Språk/Language: engelska/English ICS: 13.060.10; 13.060.70
SS-EN ISO 10253:2016
Vattenundersökningar – Bestämning av tillväxthämning på marina alger, Skeletonema arter och Phaeodactylum
tricornutum (ISO 10253:2016)
Water quality – Marine algal growth inhibition test with Skeletonema sp. and Phaeodactylum tricornutum (ISO 10253:2016)
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Europastandarden EN ISO 10253:2016 gäller som svensk standard. Detta dokument innehåller den officiella engelska versionen av EN ISO 10253:2016.
Denna standard ersätter SS-EN ISO 10253:2006, utgåva 2.
The European Standard EN ISO 10253:2016 has the status of a Swedish Standard. This document contains the official English version of EN ISO 10253:2016.
This standard supersedes the Swedish Standard SS-EN ISO 10253:2006, edition 2.
Denna standard är framtagen av kommittén för Biologiska vattenundersökningar, SIS / TK 426.
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EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN ISO 10253
November 2016
ICS 13.060.70 Supersedes EN ISO 10253:2006
English Version
Water quality - Marine algal growth inhibition test with Skeletonema sp. and Phaeodactylum tricornutum (ISO
10253:2016)
Qualité de l'eau - Essai d'inhibition de la croissance des algues marines avec Skeletonema sp. et Phaeodactylum
tricornutum (ISO 10253:2016)
Wasserbeschaffenheit - Wachstumshemmtest mit marinen Algen Skeletonema sp. und Phaeodactylum
tricornutum (ISO 10253:2016)
This European Standard was approved by CEN on 22 October 2016.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION C O M I T É E UR O P É E N DE N O R M A L I SA T I O N E UR O P Ä I SC H E S KO M I T E E F ÜR N O R M UN G
CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2016 CEN All rights of exploitation in any form and by any means reserved
worldwide for CEN national Members. Ref. No. EN ISO 10253:2016 E
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1 Scope
...1
2 Normative references
...1
3 Terms and definitions
...1
4 Principle
...2
5 Materials
...3
5.1 Test organisms
...3
5.2 Reagents
...4
6 Apparatus
...5
7 Procedure
...6
7.1 Preparation of growth medium
...6
7.2 Preparation of pre-culture and inoculum
...6
7.3 Choice of test concentrations
...6
7.4 Preparation of test substance stock solutions
...6
7.5 Preparation of test and control batches
...7
7.6 Incubation
...7
7.7 Measurements
...7
8 Validity criteria
...8
9 Interpretation of data
...8
9.1 Plotting growth curves
...8
9.2 Calculation of percentage inhibition
...8
9.3 Determination of EC(r)
x ...910 Expression of results
...9
11 Interpretation of results
...9
12 Test report
...9
Annex A (informative) Preparation of dilution series of mixtures in sea water(effluents or elutriates)
...11
Annex B (informative) Test procedure starting from stored algal inocula, and with direct measurement of algal growth in spectrophotometric cells
...12
Annex C (informative) Performance data
...18
Bibliography
...19
iii
Contents Page
European foreword ...
SS-EN ISO 10253:2016 (E)
European foreword
This document (EN ISO 10253:2016) has been prepared by Technical Committee ISO/TC 147 “Water quality” in collaboration with Technical Committee CEN/TC 230 “Water analysis” the secretariat of which is held by DIN.
This European Standard shall be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by May 2017, and conflicting national standards shall be withdrawn at the latest by May 2017.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
This document supersedes EN ISO 10253:2006.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
Endorsement notice
The text of ISO 10253:2016 has been approved by CEN as EN ISO 10253:2016 without any modification.
SS-EN ISO 10253:2016 (E)
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Water quality — Marine algal growth inhibition test with Skeletonema sp. and Phaeodactylum tricornutum
WARNING — Persons using this document should be familiar with normal laboratory practice.
This document does not purport to address all of the safety problems, if any, associated with its use. It is the responsibility of the user to establish appropriate safety and health practices and to ensure compliance with any national regulatory conditions.
IMPORTANT — It is absolutely essential that tests conducted according to this document be carried out by suitably trained staff.
1 Scope
This document specifies a method for the determination of the inhibition of growth of the unicellular marine algae Skeletonema sp. and Phaeodactylum tricornutum by substances and mixtures contained in sea water or by environmental water samples (effluents, elutriates, etc.).
The method can be used for testing substances that are readily soluble in water and are not significantly degraded or eliminated in any other way from the test medium.
NOTE With modifications, as described in ISO 14442 and ISO 5667–16, the inhibitory effects of poorly soluble organic and inorganic materials, volatile compounds, metal compounds, effluents, marine water samples and elutriates of sediments can be tested.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content constitutes requirements of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.
ISO 5667-16, Water quality — Sampling — Part 16: Guidance on biotesting of samples
ISO 14442, Water quality — Guidelines for algal growth inhibition tests with poorly soluble materials, volatile compounds, metals and waste water
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— IEC Electropedia: available at http://www.electropedia.org/
— ISO Online browsing platform: available at http://www.iso.org/obp 3.1 cell density
number of cells per unit volume of medium
Note 1 to entry: The cell density is expressed as x cells/ml.
1 SS-EN ISO 10253:2016 (E)
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3.2 specific growth rate
µ proportional rate of increase in cell density per unit of time:
µ = × 1 x 1
x t d
d ( / day ) where
x is the cell density, expressed in cells per millilitre;
t is the time, expressed in days.
Note 1 to entry: Specific growth rate is expressed in inverse days (day
−1).
3.3 growth medium
mixture of sea water and nutrients which is used for pre-cultures and controls 3.4 test medium
mixture of sea water, nutrients [growth medium (3.3)] and test material in which algal cells are incubated
3.5 test batch
mixture of sea water, nutrients and test material [test medium (3.4)] inoculated with algae 3.6 control
mixture of sea water, nutrients [growth medium (3.3)] without test material, inoculated with algae 3.7 effective concentration
EC(r)
xconcentration of test substance which results in an x % reduction in specific growth rate relative to the controls
4 Principle
Mono-specific algal strains are cultured for several generations in a defined medium containing a range of concentrations of the test substance, prepared by mixing appropriate quantities of nutrient concentrate, sea water, stock solutions of the test substance, and an inoculum of exponentially growing algal cells. The test solutions are incubated for a period of (72 ± 2) h, during which the cell density in each is measured at intervals of at least every (24 ± 2) h. Inhibition is measured as a reduction in specific growth rate, relative to control cultures grown under identical conditions.
2
SS-EN ISO 10253:2016 (E)
5 Materials
5.1 Test organisms
Use either of the following marine algae:
a) Skeletonema sp.
1)(CCAP 1077/1C, NIVA BAC 1); or
b) Phaeodactylum tricornutum Bohlin (CCAP 1052/1A, SAG 1090-1a, NIVA BAC 2).
These algae are important and widely distributed phytoplankton species (phylum Bacillariophyta) in estuarine and coastal areas.
The recommended algae are available in unialgal, non-axenic cultures from the following sources.
NIVA
Norwegian Institute for Water Research Gaustadaléen 21
N 0349 Oslo Norway CCAP
Dunstaffnage Marine Laboratory P O Box 3 Oban
Argyll PA37 1QA United Kingdom SAG
Collection of Algal Cultures University of Göttingen
Albrecht-von-Haller Institute for Plant Science Untere Karspüle 2
37073 Göttingen Germany
Stock cultures may be maintained in the medium described in 7.1. Regular subculturing is necessary.
Weekly intervals may be necessary for Skeletonema sp., every two or three weeks may be sufficient for Phaeodactylum tricornutum. The stock cultures may also be maintained for extended periods on richer algal media such as those recommended by the culture collection. It is recommended to keep the
1) The previous editions of this document suggested the use of two strains of Skeletonema costatum. Following a taxonomic review of the Skeletonema genus, several strains originally identified as S. costatum may in fact be other species. In light of this and to enable continuity in the use of previously accepted strains, the present revision of this document has changed the reference from Skeletonema costatum to Skeletonema sp. to avoid non-compliance for labs that may be using different strains.
3 SS-EN ISO 10253:2016 (E)
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