ISO 14380:2011
(Main)Water quality — Determination of the acute toxicity to Thamnocephalus platyurus (Crustacea, Anostraca)
Water quality — Determination of the acute toxicity to Thamnocephalus platyurus (Crustacea, Anostraca)
ISO 14380:2011 specifies a method for the determination of the lethal effects of toxicants to Thamnocephalus platyurus test organisms after 24 h exposure. A second method (rapid test) is described in an annex for the determination of sublethal effects after a very short exposure time (1 h). The methods are applicable to: a) chemical substances which are soluble or which can be maintained as stable suspensions or dispersions under the conditions of the test; b) industrial or sewage effluents, treated or untreated, if appropriate after decantation, filtration or centrifugation; c) fresh waters; d) aqueous extracts; e) toxins of blue‑green algae. ISO 14380:2011 is not applicable to the testing of unstable chemicals (hydrolysing, absorbing, etc.) in water unless exposure concentration is measured, nor to the testing of aquatic samples from the estuarine or marine environment.
Qualité de l'eau — Détermination de la toxicité aiguë envers Thamnocephalus platyurus (Crustacea, Anostraca)
Kakovost vode - Določevanje akutne strupenosti z raki Thamnocephalus platyurus (Crustacea, Anostraca)
Ta mednarodni standard določa metodo za določevanje smrtnih učinkov strupenih snovi na preskusne organizme Thamnocephalus platyurus po 24-urni izpostavljenosti. Druga metoda (hitri preskus) za določevanje subletalnih učinkov po zelo kratki izpostavljenosti (1 h) je opisana v dodatku A. Metode se uporabljajo za: a) kemične snovi, ki so topne ali se lahko pod pogoji preskusa ohranijo v stabilni suspenziji ali disperziji; b) obdelane ali neobdelane industrijske ali komunalne odplake, če so primerne po dekantaciji, filtriranju ali centrifugiranju; e) toksine modrozelenih alg. Ta mednarodni standard se ne uporablja za preskušanje nestabilnih kemikalij (hidrolizo, absorpcijo itd.) v vodi, razen če je bila izmerjena koncentracija izpostavljenosti, in preskušanje vodnih vzorcev iz okolja rečnih ustij ali morskega okolja.
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-junij-2012
.DNRYRVWYRGH'RORþHYDQMHDNXWQHVWUXSHQRVWL]UDNL7KDPQRFHSKDOXVSODW\XUXV
&UXVWDFHD$QRVWUDFD
Water quality - Determination of the acute toxicity to Thamnocephalus platyurus
(Crustacea, Anostraca)
Qualité de l'eau - Détermination de la toxicité aiguë envers Thamnocephalus platyurus
(Crustacea, Anostraca)
Ta slovenski standard je istoveten z: ISO 14380:2011
ICS:
13.060.70 Preiskava bioloških lastnosti Examination of biological
vode properties of water
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
INTERNATIONAL ISO
STANDARD 14380
First edition
2011-11-15
Water quality — Determination of the
acute toxicity to Thamnocephalus
platyurus (Crustacea, Anostraca)
Qualité de l’eau — Détermination de la toxicité aiguë envers
Thamnocephalus platyurus (Crustacea, Anostraca)
Reference number
©
ISO 2011
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or ISO’s
member body in the country of the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2011 – All rights reserved
Contents Page
Foreword . iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 2
5 Test environment . 2
6 Reagents, test organisms and media . 2
7 Apparatus and materials . 3
8 Treatment and preparation of samples . 4
8.1 Special precautions . 4
8.2 Preparation of the stock solutions of substances to be tested . 4
9 Procedure . 4
9.1 Hatching of the cysts . 4
9.2 Selection of test concentrations . 5
9.3 Preparation of the test and control solutions . 5
9.4 Introduction of the organisms . 6
9.5 Incubation of the test system . 6
9.6 Measurements . 7
10 Estimation of the LC . 7
11 Reference test . 8
12 Validity criteria . 8
13 Test report . 8
Annex A (informative) Rapid test for determination of sublethal effects on Thamnocephalus platyurus
(1 h exposure) .10
Annex B (informative) Culturing of Thamnocephalus platyurus for cyst production .16
Annex C (informative) Precision data.19
Bibliography .20
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO
technical committees. Each member body interested in a subject for which a technical committee has been
established has the right to be represented on that committee. International organizations, governmental and
non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International
Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 14380 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biological
methods.
iv © ISO 2011 – All rights reserved
Introduction
The evaluation of harmful effects on water quality has for several years involved the performance of biological
tests. Crustaceans are of interest from the ecotoxicological point of view because they are primary consumers
and a major component of the zooplankton in aquatic ecosystems.
The test specified in this International Standard involves determination of the lethal effects on the fresh water
fairy shrimp Thamnocephalus platyurus after 24 h exposure to the toxicant. A rapid test can also be carried out
to determine sublethal effects after a very short exposure time (1 h).
The beavertail fairy shrimp T. platyurus is to date already used extensively in toxicity testing for several reasons:
a) this anostracan crustacean has a sensitivity to chemicals which is quite similar to that of the cladoceran
crustacean Daphnia magna (see References [4][5][6][7]);
b) the assays are performed with neonates hatched from dormant eggs (cysts), which bypasses the need for
culturing or maintaining live stock cultures of test organisms;
c) T. platyurus neonates are substantially smaller than neonates of Daphnia magna, hence the assays require
much smaller test containers, and much less bench space and incubation space;
d) T. platyurus is very sensitive to cyanotoxins produced by algal blooms in eutrophicated waters (see
References [8][9]).
INTERNATIONAL STANDARD ISO 14380:2011(E)
Water quality — Determination of the acute toxicity to
Thamnocephalus platyurus (Crustacea, Anostraca)
WARNING — Persons using this International Standard should be familiar with normal laboratory
practice. This standard 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 in accordance with this International
Standard be carried out by suitably qualified staff.
1 Scope
This International Standard specifies a method for the determination of the lethal effects of toxicants to
Thamnocephalus platyurus test organisms after 24 h exposure. A second method (rapid test) is described in
Annex A for the determination of sublethal effects after a very short exposure time (1 h).
The methods are applicable to:
a) chemical substances which are soluble or which can be maintained as stable suspensions or dispersions
under the conditions of the test;
b) industrial or sewage effluents, treated or untreated, if appropriate after decantation, filtration or
centrifugation;
c) fresh waters;
d) aqueous extracts;
e) toxins of blue-green algae.
This International Standard is not applicable to the testing of unstable chemicals (hydrolysing, absorbing, etc.)
in water unless exposure concentration is measured, nor to the testing of aquatic samples from the estuarine
or marine environment.
2 Normative references
The following referenced documents are indispensable for the application 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 5814, Water quality — Determination of dissolved oxygen — Electrochemical probe method
ISO 10523, Water quality — Determination of pH
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
control batch
series of replicates containing control solution
NOTE The role of a control batch in an experimental procedure is to demonstrate the response to the detection
system imposed collectively by compounds of the matrix used in the determination, in the absence of the subject of
interest.
3.2
LC
concentration or dilution of the test sample which gives rise to 50 % mortality of the test organisms
3.3
EC
concentration or dilution of the sample which gives rise to 50 % effect on the test organisms
3.4
neonate
newly hatched individual
3.5
test batch
series of replicates filled with the same test solution
4 Principle
Freshly hatched T. platyurus larvae are exposed to a range of concentrations of the sample under analysis and
the percentage mortality of the test organisms is determined after 24 h exposure, with subsequent calculation
of the 24 h LC .
The test is carried out in one or two stages:
— a “range-finding test” to determine the range of concentrations or dilutions needed for calculation of the
24 h LC ;
— a “definitive test” conducted when the data of the range-finding test are not sufficient or adequate for
calculation of the 24 h LC .
5 Test environment
The test shall be carried out in the dark, in a temperature-controlled room or incubator at (25 ± 1) °C in the test
containers.
Maintain the atmosphere free from toxic dusts or vapours. The use of control solutions is a double check that
the test is performed in an atmosphere free from toxic dusts and vapours.
6 Reagents, test organisms and media
Use only reagents of recognized analytical grade, unless otherwise specified.
6.1 Test organisms. The test organisms are neonates of the beavertail fairy shrimp T. platyurus, which are
hatched from dormant eggs (cysts) of this crustacean.
Cysts of T. platyurus are obtained from laboratory cultures of the crustacean as described in Annex B or can
1)
be purchased from a specialized company .
1) MicroBioTests Inc., Mariakerke, Belgium, is an example of a supplier able to provide suitable Thamnocephalus platyurus
cysts commercially. This information is given for the convenience of the users of this document and does not constitute an
endorsement by ISO of this supplier.
2 © ISO 2011 – All rights reserved
6.2 Pure water, conductivity below 10 µS/cm.
6.3 Test medium, prepared by dissolving the following mineral substances in 1 l of pure water (6.2):
NaHCO 96 mg
CaSO⋅2H O 60 mg
4 2
MgSO 60 mg
KCl 4 mg
This test medium corresponds to a synthetic water of moderate hardness, i.e. containing CaCO at concentrations
of 80 mg/l to 100 mg/l (see Reference [13]). Thus prepared, the medium has a pH of 7,6 ± 0,3.
When stored in a refrigerator at (4 ± 2) °C in the dark, the solution can be used for several months.
Aerate the test medium until the dissolved oxygen concentration has reached the air saturation value and until
the pH has stabilized. If necessary, adjust the pH to 7
...
INTERNATIONAL ISO
STANDARD 14380
First edition
2011-11-15
Water quality — Determination of the
acute toxicity to Thamnocephalus
platyurus (Crustacea, Anostraca)
Qualité de l’eau — Détermination de la toxicité aiguë envers
Thamnocephalus platyurus (Crustacea, Anostraca)
Reference number
©
ISO 2011
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or ISO’s
member body in the country of the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2011 – All rights reserved
Contents Page
Foreword . iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 2
5 Test environment . 2
6 Reagents, test organisms and media . 2
7 Apparatus and materials . 3
8 Treatment and preparation of samples . 4
8.1 Special precautions . 4
8.2 Preparation of the stock solutions of substances to be tested . 4
9 Procedure . 4
9.1 Hatching of the cysts . 4
9.2 Selection of test concentrations . 5
9.3 Preparation of the test and control solutions . 5
9.4 Introduction of the organisms . 6
9.5 Incubation of the test system . 6
9.6 Measurements . 7
10 Estimation of the LC . 7
11 Reference test . 8
12 Validity criteria . 8
13 Test report . 8
Annex A (informative) Rapid test for determination of sublethal effects on Thamnocephalus platyurus
(1 h exposure) .10
Annex B (informative) Culturing of Thamnocephalus platyurus for cyst production .16
Annex C (informative) Precision data.19
Bibliography .20
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO
technical committees. Each member body interested in a subject for which a technical committee has been
established has the right to be represented on that committee. International organizations, governmental and
non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International
Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 14380 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biological
methods.
iv © ISO 2011 – All rights reserved
Introduction
The evaluation of harmful effects on water quality has for several years involved the performance of biological
tests. Crustaceans are of interest from the ecotoxicological point of view because they are primary consumers
and a major component of the zooplankton in aquatic ecosystems.
The test specified in this International Standard involves determination of the lethal effects on the fresh water
fairy shrimp Thamnocephalus platyurus after 24 h exposure to the toxicant. A rapid test can also be carried out
to determine sublethal effects after a very short exposure time (1 h).
The beavertail fairy shrimp T. platyurus is to date already used extensively in toxicity testing for several reasons:
a) this anostracan crustacean has a sensitivity to chemicals which is quite similar to that of the cladoceran
crustacean Daphnia magna (see References [4][5][6][7]);
b) the assays are performed with neonates hatched from dormant eggs (cysts), which bypasses the need for
culturing or maintaining live stock cultures of test organisms;
c) T. platyurus neonates are substantially smaller than neonates of Daphnia magna, hence the assays require
much smaller test containers, and much less bench space and incubation space;
d) T. platyurus is very sensitive to cyanotoxins produced by algal blooms in eutrophicated waters (see
References [8][9]).
INTERNATIONAL STANDARD ISO 14380:2011(E)
Water quality — Determination of the acute toxicity to
Thamnocephalus platyurus (Crustacea, Anostraca)
WARNING — Persons using this International Standard should be familiar with normal laboratory
practice. This standard 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 in accordance with this International
Standard be carried out by suitably qualified staff.
1 Scope
This International Standard specifies a method for the determination of the lethal effects of toxicants to
Thamnocephalus platyurus test organisms after 24 h exposure. A second method (rapid test) is described in
Annex A for the determination of sublethal effects after a very short exposure time (1 h).
The methods are applicable to:
a) chemical substances which are soluble or which can be maintained as stable suspensions or dispersions
under the conditions of the test;
b) industrial or sewage effluents, treated or untreated, if appropriate after decantation, filtration or
centrifugation;
c) fresh waters;
d) aqueous extracts;
e) toxins of blue-green algae.
This International Standard is not applicable to the testing of unstable chemicals (hydrolysing, absorbing, etc.)
in water unless exposure concentration is measured, nor to the testing of aquatic samples from the estuarine
or marine environment.
2 Normative references
The following referenced documents are indispensable for the application 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 5814, Water quality — Determination of dissolved oxygen — Electrochemical probe method
ISO 10523, Water quality — Determination of pH
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
control batch
series of replicates containing control solution
NOTE The role of a control batch in an experimental procedure is to demonstrate the response to the detection
system imposed collectively by compounds of the matrix used in the determination, in the absence of the subject of
interest.
3.2
LC
concentration or dilution of the test sample which gives rise to 50 % mortality of the test organisms
3.3
EC
concentration or dilution of the sample which gives rise to 50 % effect on the test organisms
3.4
neonate
newly hatched individual
3.5
test batch
series of replicates filled with the same test solution
4 Principle
Freshly hatched T. platyurus larvae are exposed to a range of concentrations of the sample under analysis and
the percentage mortality of the test organisms is determined after 24 h exposure, with subsequent calculation
of the 24 h LC .
The test is carried out in one or two stages:
— a “range-finding test” to determine the range of concentrations or dilutions needed for calculation of the
24 h LC ;
— a “definitive test” conducted when the data of the range-finding test are not sufficient or adequate for
calculation of the 24 h LC .
5 Test environment
The test shall be carried out in the dark, in a temperature-controlled room or incubator at (25 ± 1) °C in the test
containers.
Maintain the atmosphere free from toxic dusts or vapours. The use of control solutions is a double check that
the test is performed in an atmosphere free from toxic dusts and vapours.
6 Reagents, test organisms and media
Use only reagents of recognized analytical grade, unless otherwise specified.
6.1 Test organisms. The test organisms are neonates of the beavertail fairy shrimp T. platyurus, which are
hatched from dormant eggs (cysts) of this crustacean.
Cysts of T. platyurus are obtained from laboratory cultures of the crustacean as described in Annex B or can
1)
be purchased from a specialized company .
1) MicroBioTests Inc., Mariakerke, Belgium, is an example of a supplier able to provide suitable Thamnocephalus platyurus
cysts commercially. This information is given for the convenience of the users of this document and does not constitute an
endorsement by ISO of this supplier.
2 © ISO 2011 – All rights reserved
6.2 Pure water, conductivity below 10 µS/cm.
6.3 Test medium, prepared by dissolving the following mineral substances in 1 l of pure water (6.2):
NaHCO 96 mg
CaSO⋅2H O 60 mg
4 2
MgSO 60 mg
KCl 4 mg
This test medium corresponds to a synthetic water of moderate hardness, i.e. containing CaCO at concentrations
of 80 mg/l to 100 mg/l (see Reference [13]). Thus prepared, the medium has a pH of 7,6 ± 0,3.
When stored in a refrigerator at (4 ± 2) °C in the dark, the solution can be used for several months.
Aerate the test medium until the dissolved oxygen concentration has reached the air saturation value and until
the pH has stabilized. If necessary, adjust the pH to 7,6 ± 0,3 using sodium hydroxide or hydrochloric acid
solutions. The concentration of the acid or base required shall be selected so that the volume to be admixed is
as small as possible. Bring the temperature of the test medium up to (25 ± 1) °C prior to use.
6.4 Hatching medium. An eightfold dilution of the test medium (6.3) with pure water (6.2).
6.5 Reference substance. Potassium dichromate (K Cr O ) is the recommended reference chemical.
2 2 7
7 Apparatus and materials
Usual laboratory apparatus and glassware and in particular the following.
7.1 Temperature-controlled room or chamber.
7.2 Hatching Petri dishes, small Petri dishes, diameter 5 cm, in glass or in inert plastic material.
7.3 Test containers. Disposable microplates made from chemically inert material, comprising wells with
a capacity >1 ml. For example, 24 (4 × 6) well microplates with a well diameter of approximately 16 mm are
suitable.
7.4 Pipette for sampling the test organisms, with a sufficient diameter for capturing the animals while
allowing sampling of o
...
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