Environmental characterization of leachates from waste and soil using reproductive and toxicological gene expression in Daphnia magna

This document specifies the crucial steps of a quantitative real-time polymerase chain reaction (qPCR) method to quantify the abundance of specific mRNA molecules extracted from Daphnia magna.
The method allows the identification of molecular responses to exposures for potentially toxic substances through the analysis of the abundance of specific mRNA molecules. In this document, the central genes involved in reproductive and toxic responses are included.
NOTE   The selection of genes can be adapted to specific exposure conditions, for example, exposure to known toxic substances, by adding genes known to respond to a specific insult.
The present method allows for rapid, robust and sensitive detection of molecular responses and can be used to analyse the toxic effects of water leachates from soil and waste. The method gives information of the concentration of a substance or test-liquid at which toxic effects begin to occur prior to observations of reproductive or toxic effects at higher levels of organization, which reduces the need for the use of safety factors in toxicity assessment.
The method is useful in several types of risk assessment. In this document, the genes studied are appropriate for the assessment of the risks when recycling materials and for the classification of waste, but the method can be adapted to other types of risk assessment by including other genes.

Umwelttechnische Charakterisierung von Sickerwässern aus Abfall und Boden mittels reproduktiver und toxikologischer Genexpression in Daphnia magna

Dieses Dokument legt die entscheidenden Schritte einer quantitativen Echtzeit-Polymerase-Kettenreaktion (qPCR) zur Quantifizierung der Anzahl spezifischer mRNA Moleküle fest, die aus Daphnia magna extrahiert wurden.
Dieses Verfahren ermöglicht durch die Analyse der Anzahl spezifischer mRNA Moleküle die Identifikation molekularer Reaktionen auf die Exposition gegenüber potenziell toxischen Stoffen. Dieses Dokument deckt auch die zentralen Gene ab, die an reproduktionsbezogenen und toxischen Reaktionen beteiligt sind.
ANMERKUNG   Die Auswahl der Gene kann an spezifische Expositionsbedingungen angepasst werden, z. B. an die Exposition gegenüber bekannten toxischen Stoffen, indem Gene hinzugefügt werden, die eine bekannte Reaktion auf einen spezifischen Einfluss haben.
Das vorliegende Verfahren ermöglicht einen schnellen, robusten und empfindlichen Nachweis molekularer Reaktionen und kann zur Analyse der toxischen Wirkungen von Sickerwasser aus Böden und Abfällen verwendet werden. Das Verfahren liefert Informationen über die Konzentration einer Substanz oder Prüfflüssigkeit, bei der erste toxische Wirkungen auftreten, bevor reproduktionsbezogene oder toxische Wirkungen auf höheren Organisationsebenen beobachtet werden, wodurch die Notwendigkeit der Verwendung von Sicherheitsfaktoren in der Toxizitätsbeurteilung verringert wird.
Das Verfahren ist für verschiedene Arten der Gefährdungseinschätzung nützlich. Die in diesem Dokument untersuchten Gene sind für die Beurteilung der Risiken beim Recycling von Stoffen und für die Klassifizierung von Abfall geeignet, das Verfahren kann durch Aufnahme anderer Gene jedoch auch an andere Arten der Gefährdungseinschätzung angepasst werden.

Caractérisation environnementale des lixiviats de déchets et de sols à l’aide de l’expression génétique reproductive et toxicologique chez Daphnia magna

Le présent document spécifie les étapes cruciales d’une méthode de réaction de polymérisation en chaîne (qPCR) quantitative en temps réel pour quantifier l’abondance de molécules d’ARNm spécifiques extraites de Daphnia magna.
La méthode permet d’identifier les réponses moléculaires aux expositions à des substances potentiellement toxiques par l’analyse de l’abondance de molécules d’ARNm spécifiques. Dans le présent document, les gènes principaux impliqués dans les réponses reproductives et toxiques sont inclus.
NOTE   La sélection de gènes peut être adaptée à des conditions d’exposition spécifiques, par exemple l’exposition à des substances toxiques connues, en ajoutant des gènes connus pour répondre à une agression spécifique.
La présente méthode permet une détection rapide, fiable et sensible des réponses moléculaires et peut être utilisée pour analyser les effets toxiques des lixiviats des sols et des déchets ainsi que ceux des eaux réceptrices. La méthode donne des informations sur la concentration d’une substance ou d’un liquide d’essai à laquelle les effets toxiques commencent à se manifester avant l’observation d’effets reproductifs ou toxiques à des niveaux d’organisation plus élevés, ce qui réduit la nécessité d’utiliser des facteurs de sécurité dans l’évaluation de la toxicité.
La présente méthode est utile dans plusieurs types d’évaluation du risque. Dans le présent document, les gènes étudiés sont appropriés pour l’évaluation du risque lors du recyclage des matériaux et pour la classification des déchets, mais la méthode peut être adaptée à d’autres types d’évaluation en incluant d’autres gènes.

Karakterizacija izlužkov odpadkov in tal z reproduktivno in toksikološko ekspresijo genov pri Daphnia magna

Ta dokument določa ključne korake pri metodi kvantitativne verižne reakcije s polimerazo v realnem času (qPCR) za kvantificiranje številčnosti specifičnih molekul mRNA, ekstrahiranih iz Daphnia magna.
Metoda omogoča identifikacijo molekularnih odzivov na izpostavljenost potencialno strupenim snovem z analizo številčnosti specifičnih molekul mRNA. V tem dokumentu so vključeni osrednji geni, vključeni v reproduktivne in toksične odzive.
OPOMBA:   Izbor genov je mogoče prilagoditi specifičnim pogojem izpostavljenosti, na primer izpostavljenosti znanim strupenim snovem, z dodajanjem genov, za katere je znano, da se odzivajo na določeno poškodbo.
Ta metoda omogoča hitro, robustno in občutljivo zaznavanje molekularnih odzivov in se lahko uporablja za analizo toksičnih učinkov vodnih izcedkov iz zemlje in odpadkov. Z metodo se dobi informacije o koncentraciji snovi ali preskusne tekočine, pri kateri se pojavijo toksični učinki, preden se na višjih organizacijskih ravneh opazijo reproduktivni ali toksični učinki, kar zmanjšuje potrebo po uporabi varnostnih faktorjev pri oceni toksičnosti.
Metoda se uporablja za različne vrste ocenjevanja tveganja. V tem dokumentu so preučevani geni primerni za oceno tveganja pri recikliranju materialov in za klasifikacijo odpadkov, vendar je mogoče metodo prilagoditi drugim vrstam ocene tveganja z vključitvijo drugih genov.

General Information

Status
Published
Public Enquiry End Date
02-Oct-2022
Publication Date
12-Feb-2023
Technical Committee
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
10-Feb-2023
Due Date
17-Apr-2023
Completion Date
13-Feb-2023

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Standards Content (Sample)

SLOVENSKI STANDARD
SIST-TS CEN/TS 17883:2023
01-marec-2023
Okoljska karakterizacija izcednih voda iz odpadkov in tal z reproduktivno in
toksikološko ekspresijo genov pri Daphnia magna
Environmental characterization of leachates from waste and soil using reproductive and
toxicological gene expression in Daphnia magna
Umwelttechnische Charakterisierung von Sickerwässern aus Abfall und Boden mittels
reproduktiver und toxikologischer Genexpression in Daphnia magna
Caractérisation environnementale des lixiviats de déchets et de sols à l’aide de
l’expression génétique reproductive et toxicologique chez Daphnia magna
Ta slovenski standard je istoveten z: CEN/TS 17883:2022
ICS:
13.030.01 Odpadki na splošno Wastes in general
13.060.70 Preiskava bioloških lastnosti Examination of biological
vode properties of water
13.080.99 Drugi standardi v zvezi s Other standards related to
kakovostjo tal soil quality
SIST-TS CEN/TS 17883:2023 en,fr
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------
SIST-TS CEN/TS 17883:2023

---------------------- Page: 2 ----------------------
SIST-TS CEN/TS 17883:2023


CEN/TS 17883
TECHNICAL SPECIFICATION

SPÉCIFICATION TECHNIQUE

December 2022
TECHNISCHE SPEZIFIKATION
ICS 13.030.01; 13.060.70; 13.080.99
English Version

Environmental characterization of leachates from waste
and soil using reproductive and toxicological gene
expression in Daphnia magna
Caractérisation environnementale des lixiviats de Umwelttechnische Charakterisierung von
déchets et de sols à l'aide de l'expression génétique Sickerwässern aus Abfall und Boden mittels
reproductive et toxicologique chez Daphnia magna reproduktiver und toxikologischer Genexpression in
Daphnia magna
This Technical Specification (CEN/TS) was approved by CEN on 14 November 2022 for provisional application.

The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to
submit their comments, particularly on the question whether the CEN/TS can be converted into a European Standard.

CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS
available promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in
parallel to the CEN/TS) until the final decision about the possible conversion of the CEN/TS into an EN is reached.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and
United Kingdom.





EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALI S A T IO N

EUROPÄISCHES KOMITEE FÜR NORMUN G

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2022 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TS 17883:2022 E
worldwide for CEN national Members.

---------------------- Page: 3 ----------------------
SIST-TS CEN/TS 17883:2023
CEN/TS 17883:2022 (E)
Contents Page
European foreword . 3
Introduction . 4
1 Scope . 8
2 Normative references . 8
3 Terms and definitions . 8
4 Principle . 9
4.1 General. 9
4.2 Toxicogenomic qPCR method . 10
4.3 Exposure of Daphnia magna . 10
4.4 Choice of genes for study . 11
5 Tes
...

SLOVENSKI STANDARD
SIST-TS CEN/TS 17883:2023
01-marec-2023
Karakterizacija izlužkov odpadkov in tal z reproduktivno in toksikološko
ekspresijo genov pri Daphnia magna
Environmental characterization of leachates from waste and soil using reproductive and
toxicological gene expression in Daphnia magna
Umwelttechnische Charakterisierung von Sickerwässern aus Abfall und Boden mittels
reproduktiver und toxikologischer Genexpression in Daphnia magna
Caractérisation environnementale des lixiviats de déchets et de sols à l’aide de
l’expression génétique reproductive et toxicologique chez Daphnia magna
Ta slovenski standard je istoveten z: CEN/TS 17883:2022
ICS:
13.030.01 Odpadki na splošno Wastes in general
13.060.70 Preiskava bioloških lastnosti Examination of biological
vode properties of water
13.080.99 Drugi standardi v zvezi s Other standards related to
kakovostjo tal soil quality
SIST-TS CEN/TS 17883:2023 en,fr
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------
SIST-TS CEN/TS 17883:2023

---------------------- Page: 2 ----------------------
SIST-TS CEN/TS 17883:2023


CEN/TS 17883
TECHNICAL SPECIFICATION

SPÉCIFICATION TECHNIQUE

December 2022
TECHNISCHE SPEZIFIKATION
ICS 13.030.01; 13.060.70; 13.080.99
English Version

Environmental characterization of leachates from waste
and soil using reproductive and toxicological gene
expression in Daphnia magna
Caractérisation environnementale des lixiviats de Umwelttechnische Charakterisierung von
déchets et de sols à l'aide de l'expression génétique Sickerwässern aus Abfall und Boden mittels
reproductive et toxicologique chez Daphnia magna reproduktiver und toxikologischer Genexpression in
Daphnia magna
This Technical Specification (CEN/TS) was approved by CEN on 14 November 2022 for provisional application.

The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to
submit their comments, particularly on the question whether the CEN/TS can be converted into a European Standard.

CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS
available promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in
parallel to the CEN/TS) until the final decision about the possible conversion of the CEN/TS into an EN is reached.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and
United Kingdom.





EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALI S A T IO N

EUROPÄISCHES KOMITEE FÜR NORMUN G

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2022 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TS 17883:2022 E
worldwide for CEN national Members.

---------------------- Page: 3 ----------------------
SIST-TS CEN/TS 17883:2023
CEN/TS 17883:2022 (E)
Contents Page
European foreword . 3
Introduction . 4
1 Scope . 8
2 Normative references . 8
3 Terms and definitions . 8
4 Principle . 9
4.1 General. 9
4.2 Toxicogenomic qPCR method . 10
4.3 Exposure of Daphnia magna . 10
4.4 Choice of genes for study . 11
5 Test materials .
...

SLOVENSKI STANDARD
kSIST-TS FprCEN/TS 17883:2022
01-september-2022
Okoljska karakterizacija izcednih voda iz odpadkov in tal z reproduktivno in
toksikološko ekspresijo genov pri Daphnia magna
Environmental characterization of leachates from waste and soil using reproductive and
toxicological gene expression in Daphnia magna
Umwelttechnische Charakterisierung von Sickerwässern aus Abfall und Boden mittels
reproduktiver und toxikologischer Genexpression in Daphnia magna
Ta slovenski standard je istoveten z: FprCEN/TS 17883
ICS:
13.030.01 Odpadki na splošno Wastes in general
13.060.70 Preiskava bioloških lastnosti Examination of biological
vode properties of water
13.080.99 Drugi standardi v zvezi s Other standards related to
kakovostjo tal soil quality
kSIST-TS FprCEN/TS 17883:2022 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------
kSIST-TS FprCEN/TS 17883:2022

---------------------- Page: 2 ----------------------
kSIST-TS FprCEN/TS 17883:2022


FINAL DRAFT
TECHNICAL SPECIFICATION
FprCEN/TS 17883
SPÉCIFICATION TECHNIQUE

TECHNISCHE SPEZIFIKATION

July 2022
ICS 13.030.01; 13.060.70; 13.080.99
English Version

Environmental characterization of leachates from waste
and soil using reproductive and toxicological gene
expression in Daphnia magna



This draft Technical Specification is submitted to CEN members for Vote. It has been drawn up by the Technical Committee
CEN/TC 444.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and
United Kingdom.

Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are
aware and to provide supporting documentation.

Warning : This document is not a Technical Specification. It is distributed for review and comments. It is subject to change
without notice and shall not be referred to as a Technical Specification.


EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATIO N

EUROPÄISCHES KOMITEE FÜR NORMUN G

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2022 CEN All rights of exploitation in any form and by any means reserved Ref. No. FprCEN/TS 17883:2022 E
worldwide for CEN national Members.

---------------------- Page: 3 ----------------------
kSIST-TS FprCEN/TS 17883:2022
FprCEN/TS 17883:2022 (E)
Contents Page
European foreword . 3
Introduction . 4
1 Scope . 9
2 Normative references . 9
3 Terms and definitions . 9
4 Principle . 10
4.1 General. 10
4.2 Toxicogenomic qPCR method . 11
4.3 Exposure of Daphnia magna . 11
4.4 Choice of genes for study . 12
5 Test materials . 13
5.1 Daphnia magna reference water . 13
5.2 Daphnia magna culture . 14
5.3 Reagents . 14
6 Apparatus .
...

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