natgillin/translations-raw huggingface dataset:
Angiv, hvilken beholder der er anvendt til forsøget
The stock animals should be maintained in culture conditions (light, temperature, medium, feeding and animals per unit volume) similar to those to be used in the test.
Registrer aborteret yngel som ‘AB’ i den relevante rubrik
If the Daphnia culture medium to be used in the test is different from that used for routine Daphnia culture, it is good practice to include a pre-test acclimation period of normally about 3 weeks (i.e. one generation) to avoid stressing the parent animals.
Registrer forældredyrs dødelighed som ‘M’ i den relevante rubrik
Test medium
EKSEMPEL PÅ DATAARK MED RESULTATER AF KEMISKE ANALYSER
It is recommended that a fully defined medium be used in this test.
Målte koncentrationer
This can avoid the use of additives (e.g. seaweed, soil extract), which are difficult to characterise, and therefore improves the opportunities for standardisation between laboratories.
Nominel konc.
Elendt M4 (6) and M7 media (see Appendix 2) have been found to be suitable for this purpose.
Prøve i uge 1
However, other media (e.g. (7) (8)) are acceptable provided the performance of the Daphnia culture is shown to meet the validity criteria for the test.
Prøve i uge 2
If media are used which include undefined additives, these additives should be specified clearly and information should be provided in the test report on composition, particularly with regard to carbon content as this may contribute to the diet provided.
Prøve i uge 3
It is recommended that the total organic carbon (TOC) and/or chemical oxygen demand (COD) of the stock preparation of the organic additive be determined and an estimate of the resulting contribution to the TOC/COD in the test medium made.
gammel
It is further recommended that TOC levels in the medium (i.e. before addition of the algae) be below 2 mg/l (9).
Målte koncentrationer i procent af nominel konc.
When testing chemicals containing metals, it is important to recognise that the properties of the test medium (e.g. hardness, chelating capacity) may have a bearing on the toxicity of the test chemical.
BEREGNING AF TIDSVÆGTET GENNEMSNIT
For this reason, a fully defined medium is desirable.
Tidsvægtet gennemsnit
However, at present, the only fully defined media which are known to be suitable for long-term culture of Daphnia magna are Elendt M4 and M7.
Da koncentrationen af testkemikaliet kan falde i perioden mellem udskiftningerne af mediet, er det nødvendigt at overveje, hvilken koncentration der skal vælges som repræsentant for variationen af koncentrationer, som den voksne dafnie bliver udsat for.
Both media contain the chelating agent EDTA.
Valget bør baseres på såvel biologiske som statistiske betragtninger.
Work has shown (2) that the ‘apparent toxicity’ of cadmium is generally lower when the reproduction test is performed in M4 and M7 media than in media containing no EDTA.
Hvis formeringen for eksempel formodes mest påvirket af den højeste koncentration, anvendes denne.
M4 and M7 are not, therefore, recommended for testing chemicals containing metals, and other media containing known chelating agents should also be avoided.
Hvis den akkumulerede virkning eller langtidsvirkningen af det toksiske kemikalie imidlertid anses for mere vigtig, er gennemsnitskoncentrationen mere relevant.
For metal-containing chemicals it may be advisable to use an alternative medium such as, for example, ASTM reconstituted hard fresh water (9), which contains no EDTA.
I dette tilfælde er det hensigtsmæssigt at anvende den tidsvægtede gennemsnitskoncentration, eftersom der her er taget hensyn til variationen i baggrundskoncentration over tid.
This combination of ASTM reconstituted hard fresh water and seaweed extract (10) is suitable for long-term culturing of Daphnia magna (2).
Eksempel på tidsvægtet gennemsnit
The dissolved oxygen concentration should be above 3 mg/l at the beginning and during the test.
Dage
The pH should be within the range 6 - 9, and normally it should not vary by more than 1,5 units in any one test.
Fig. 1 viser et eksempel på et (forenklet) forsøg over syv dage med udskiftning af mediet på dag 0, 2 og 4.
Hardness above 140 mg/l (as CaCO3) is recommended.
Den tynde zigzaglinje repræsenterer koncentrationen på ethvert tidspunkt.
Tests at this level and above have demonstrated reproductive performance in compliance with the validity criteria (11) (12).
Faldet i koncentration formodes at følge en eksponential henfaldsproces.
Test solutions of the chosen concentrations are usually prepared by dilution of a stock solution.
De seks angivne punkter repræsenterer de målte koncentrationer ved begyndelsen og slutningen af hver udskiftningsperiode.
Stock solutions should preferably be prepared, without using any solvents or dispersants if possible, by mixing or agitating the test chemical in test medium using mechanical means such as agitating, stirring or ultrasonication, or other appropriate methods.
Den tykke, solide linje angiver placeringen af det tidsvægtede gennemsnit.
It is preferable to expose test systems to concentrations of the test chemical to be used in the study for as long as is required to demonstrate the maintenance of stable exposure concentrations prior to the introduction of test organisms.
Det tidsvægtede gennemsnit er beregnet, så arealet under det tidsvægtede gennemsnit har samme størrelse som arealet under koncentrationskurven.
If the test chemical is difficult to dissolve in water, procedures described in the OECD Guidance for handling difficult substances should be followed (13).
Beregningen af det ovenfor viste eksempel ses i tabel 1.
The use of solvents or dispersants should be avoided, but may be necessary in some cases in order to produce a suitably concentrated stock solution for dosing.
Beregning af tidsvægtet gennemsnit
A dilution water control with adequate replicates and, if unavoidable, a solvent control with adequate replicates should be run in addition to the test concentrations.
Udskiftning Antal
Only solvents or dispersants that have been investigated to have no significant or only minimal effects on the response variable should be used in the test.
Conc 0
Examples of suitable solvents (e.g. acetone, ethanol, methanol, dimethylformamide and triethylene glycol) and dispersants (e.g. Cremophor RH40, methylcellulose 0,01 % and HCO-40) are given in (13).
Areal
Where a solvent or dispersant is used, its final concentration should not be greater than 0,1 ml/l (13) and it should be the same concentration in all test vessels, except the dilution water control.
Antal dage i alt
However, every effort should be made to keep the solvent concentration to a minimum.
Ln(Conc 0) er den naturlige logaritme til Conc 0.
For semi-static tests, at least 10 animals individually held at each test concentration and at least 10 animals individually held in the control series.
Ln(Conc 1) er den naturlige logaritme til Conc 1.
For flow-through tests, 40 animals divided into four groups of 10 animals at each test concentration has been shown to be suitable (1).
Arealet er arealet under eksponentialkurven for hver udskiftningsperiode.
A smaller number of test organisms may be used and a minimum of 20 animals per concentration divided into two or more replicates with an equal number of animals (e.g. four replicates each with five daphnids) is recommended.
Det beregnes således:
Note that for tests where animals are held in groups, it will not be possible to exclude any offspring from the statistical analysis if inadvertent/ accidental parental mortality occurs when the reproduction has begun, and hence in these cases the reproductive output should be expressed as total number of living offspring produced per parent present at the beginning of the test.
Det tidsvægtede gennemsnit er det totale areal divideret med det totale antal dage.
Treatments should be allocated to the test vessels and all subsequent handling of the test vessels should be done in a random fashion.
Tabellen vil selvfølgelig skulle forlænges til 21 dage i forbindelse med dafnie-formeringstesten.
Failure to do this may result in bias that could be construed as being a concentration effect.
Det er klart, at når der kun foretages målinger ved begyndelsen og slutningen af hver udskiftningsperiode, kan man ikke bekræfte, at henfaldsprocessen er eksponentiel.
In particular, if experimental units are handled in treatment or concentration order, then some time-related effect, such as operator fatigue or other error, could lead to greater effects at the higher concentrations.
En anderledes kurve ville resultere i en anderledes beregning for arealet.
Furthermore, if the test results are likely to be affected by an initial or environmental condition of the test, such as position in the laboratory, then consideration should be given to blocking the test.
En eksponential henfaldsproces er dog ikke usandsynlig, og det er den bedste kurve at anvende, når der ikke foreligger andre oplysninger.
For semi-static tests, feeding should preferably be done daily, but at least three times per week (i.e. corresponding to media changes).
Der må dog udvises stor omhu, hvis en kemisk analyse ikke kan påvise et testkemikalie ved afslutningen af udskiftningsperioden.
The possible dilution of the exposure concentrations by food addition should be taken into account and avoided as much as possible with well concentrated algae suspensions.
Medmindre man kan påvise, med hvilken hastighed testkemikaliet forsvandt fra opløsningen, er det ikke muligt at finde frem til et realistisk areal under kurven, hvilket medfører, at det er umuligt at finde frem til et rimeligt tidsvægtet gennemsnit.
Deviations from this (e.g. for flow-through tests) should be reported.
VEJLEDNING TIL IDENTIFIKATION AF NYFØDTES KØN
During the test, the diet of the parent animals should preferably be living algal cells of one or more of the following: Chlorella sp., Pseudokirchneriella subcapitata (formerly Selenastrum capricornutum) and Desmodesmus subspicatus (formerly Scenedesmus subspicatus).
Produktion af nyfødte hanner kan forekomme under skiftende miljøforhold, f.eks. ved forkortet lysperiode, temperaturændring, faldende foderkoncentration og øget populationstæthed (Hobæk og Larson, 1990, Kleiven et al. 1992).
The supplied diet should be based on the amount of organic carbon (C) provided to each parent animal.
Produktion af hanner er også en kendt respons på visse midler til regulering af insektvækst (Oda et al., 2005).
Research (14) has shown that, for Daphnia magna, ration levels of between 0,1 and 0,2 mg C/Daphnia/day are sufficient for achieving the required number of living offspring to meet the test validity criteria.
Under forhold, hvor kemiske stressfaktorer medfører en nedgang i antallet af formeringsdygtigt afkom fra parthenogenetiske hunner, ville man forvente et stigende antal hanner (OECD, 2008).
The ration can be supplied either at a constant rate throughout the period of the test, or, if desired, a lower rate can be used at the beginning and then increased during the test to take account of growth of the parent animals.
På grundlag af de foreliggende oplysninger er det ikke muligt at forudsige, hvad der er mest følsomt, kønsfordelingen eller reproduktionsendepunktet.
In this case, the ration should still remain within the recommended range of 0,1 - 0,2 mg C/Daphnia/day at all times.
Der er dog indikationer (se del 1 i valideringsrapporten) for, at denne stigning i antallet af hanner kan være mindre følsom end nedgangen i afkom.
If surrogate measures, such as algal cell number or light absorbance, are to be used to feed the required ration level (i.e. for convenience since measurement of carbon content is time consuming), each laboratory should produce its own nomograph relating the surrogate measure to carbon content of the algal culture (see Appendix 3 for advice on nomograph production).
Da det primære formål med forsøgsmetoden er at vurdere antallet af afkom, er forekomsten af hanner en valgfri observation.
Nomographs should be checked at least annually and more frequently if algal culture conditions have changed.
Hvis dette valgfri endepunkt evalueres i en undersøgelse, bør et supplerende validitetskriterium i testen være højst 5 % hanner.
Light absorbance has been found to be a better surrogate for carbon content than cell number (15).
Den mest praktiske og letteste måde at inddele dafnierne efter køn på er at bruge deres fænotypiske egenskaber, da hanner og hunner er genetisk identiske og deres køn miljømæssigt bestemt.
A concentrated algal suspension should be fed to the Daphnia to minimise the volume of algal culture medium transferred to the test vessels.
Hanner og hunner er forskellige, hvad angår de første antenners længde og morfologi, idet hannernes antenner er længere end hunnernes (fig. 1).
Concentration of the algae can be achieved by centrifugation followed by re-suspension in Daphnia culture medium.
Denne forskel kan observeres umiddelbart efter fødslen, selv om andre sekundære kønskarakteristika udvikler sig i takt med deres vækst (se f.eks. fig. 2 i Olmstead and LeBlanc, 2000).
Light
Mediet holdes på et minimum for at begrænse dyrenes bevægelsesfrihed.
For light-measuring instruments calibrated in lux, an equivalent range of 1000-1500 lux for cool white light corresponds close to the recommended light intensity 15-20 μE · m-2 · s-1.
De første antenner kan observeres under et stereomikroskop (× 10-60).
The temperature of the test media should be within the range 18-22 °C.
24 timer gammel han (til venstre) og hun (til højre) af D. magna.
However, for any one test, the temperature should not, if possible, vary by more than 2 °C within these limits (e.g. 18-20, 19-21 or 20-22 °C) as daily range.
Hanner kan skelnes fra hunner ved de første antenners længde og morfologi som vist i cirklerne (Tatarazako et al., 2004).
It may be appropriate to use an additional test vessel for the purposes of temperature monitoring.
HENVISNINGER
Aeration
I del C affattes kapitel C.29, punkt 66 som følger:
The test vessels should not be aerated during the test.
En test anses for gyldig, hvis:
Test design
den gennemsnitlige procentvise nedbrydning i FC-beholdere, som indeholder referencekemikaliet, er > 60 % på den 14. inkubationsdag, og
Range finding test
den gennemsnitlige mængde af TIC i blindkontrollerne FB ved slutningen af testen er < 3 mg C/l.
When necessary, a range-finding test is conducted with, for example five test chemical concentrations and two replicates for each treatment and control.
Hvis f.eks. en høj blindproduktion af uorganisk kulstof er et problem, følges fremgangsmåden i afsnit 27-32.«
The duration of the range-finding test is 21 days or of a sufficient duration to reliably predict effect levels.
I del C indsættes følgende kapitler:
At the end of the test, reproduction of the Daphnia is assessed.
Toksicitetstest med fisk i de tidlige udviklingsstadier
The number of parents and the occurrence of offspring should be recorded.
Denne forsøgsmetode svarer til OECD-testvejledning (TG) 210 (2013).
Definitive test
Test med fisk i de tidlige udviklingsstadier har til formål at bestemme kemikaliers letale og subletale virkninger på de testede stadier og arter.
Normally there should be at least five test concentrations, bracketing effective concentration (e.g. ECx), and arranged in a geometric series with a separation factor preferably not exceeding 3,2 An appropriate number of replicates for each test concentration should be used (see paragraphs 24-25).
De giver nyttige oplysninger til estimering af kemikaliets kroniske letale og subletale virkninger på andre fiskearter.
Justification should be provided if fewer than five concentrations are used.
Testvejledning 210 er baseret på et forslag fra Det Forenede Kongerige, som blev drøftet på et møde med deltagelse af OECD-eksperter i Medmenham (Det Forenede Kongerige) i november 1988, og blev opdateret i 2013 for at afspejle erfaringerne med brug af testen og anbefalinger fra en OECD-workshop om toksicitetstest med fisk, som blev afholdt i september 2010 (1).
Chemicals should not be tested above their solubility limit in test medium.
Fisk i de tidlige udviklingsstadier udsættes for en række koncentrationer af testkemikaliet opløst i vand.
Before conducting the experiment it is advisable to consider the statistical power of the tests design and using appropriate statistical methods (4).
Gennemstrømningsbetingelser foretrækkes, men hvis det ikke kan lade sig gøre, er semistatiske betingelser acceptable.
In setting the range of concentrations, the following should be borne in mind:
Nærmere oplysninger findes i OECD Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures (2).
When ECx for effects on reproduction is estimated, it is advisable that sufficient concentrations are used to define the ECx with an appropriate level of confidence.
Testen indledes ved at anbringe befrugtede æg i testkamre og fortsættes i det artsspecifikke tidsrum, der er nødvendigt for, at kontrolfiskene når yngelstadiet.
Test concentrations used should preferably bracket the estimated ECx such that ECx is found by interpolation rather than extrapolation.
Letale og subletale virkninger måles og sammenlignes med kontrolværdierne for at bestemme den laveste koncentration med observeret effekt (LOEC) med henblik på at bestemme i) koncentrationen uden observeret effekt (NOEC) og/eller ii) ECx (f.eks. EC10, EC20) ved hjælp af en regressionsmodel for at beregne den koncentration, som ville forårsage en ændring i den målte virkning på x %.
It is an advantage for the following statistical analysis to have more test concentrations (e.g. 10) and fewer replicates of each concentration (e.g. 5 thus holding the total number of vessels constant) and with 10 controls.
Rapporteringen af relevante effektkoncentrationer og parametre kan afhænge af de lovgivningsmæssige rammer.
When estimating the LOEC and/or NOEC, the lowest test concentration should be low enough so that the reproductive output at that concentration is not significantly lower than that in the control.
ECx skal ligge i det område, testkoncentrationerne dækker, således at ECx opnås ved interpolering og ikke ved ekstrapolering (definitioner er anført i tillæg 1).
If this is not the case, the test should be repeated with a reduced lowest concentration.
Ved testkemikaliet forstås det kemikalie, der testes.
When estimating the LOEC and/or NOEC, the highest test concentration should be high enough so that the reproductive output at that concentration is significantly lower than that in the control.
Testkemikaliets vandopløselighed (se kapitel A.6 i dette bilag) og damptryk (se kapitel A.4 i dette bilag) bør være kendt, og der bør foreligge en pålidelig analysemetode til kvantificering af kemikaliet i testopløsningerne med kendt og publiceret nøjagtighed og nedre målegrænse.
If this is not the case, the test should be repeated with an increased highest concentration unless the maximum required test concentration for chronic effects testing (i.e., 10 mg/l) was used as the highest test concentration in the initial test.
Selv om det ikke er nødvendigt for at gennemføre testen, kan resultater fra en akut toksicitetstest (se kapitel C.1 eller C.49 i dette bilag), fortrinsvis udført på den art, der er valgt til denne test, give nyttige oplysninger.
If no effects are observed at the highest concentration in the range-finding test (e.g. at 10 mg/l), or when the test chemical is highly likely to be of low/ no toxicity based on lack of toxicity to other organisms and/or low/no uptake, the reproduction test may be performed as a limit test, using a test concentration of e.g.10 mg/l and the control.
Hvis forsøgsmetoden anvendes til at teste en blanding, skal der så vidt muligt foreligge oplysninger om dens sammensætning, f.eks. i form af bestanddelenes kemiske identitet, deres kvantitative forekomst og deres stofspecifikke egenskaber (som de ovenfor nævnte).
Ten replicates should be used for both the treatment and the control groups.
Før brug af forsøgsmetoden til lovmæssig test af en blanding bør det overvejes, om den vil give resultater, som er egnede til det lovgivningsmæssige anvendelsesformål.
When a limit test might need to be done in a flow-through system less replicates would be adequate.
Nyttige oplysninger omfatter strukturformlen, stoffets renhed, vandopløselighed, vand- og lysstabilitet, pKa, Pow samt resultater fra en test for umiddelbar biologisk nedbrydelighed (f.eks. kapitel C.4 eller C.29 i dette bilag).
A limit test will provide the opportunity to demonstrate that there is no statistically significant effect at the limit concentration, but if effects are recorded a full test will normally be required.
koncentrationen af opløst oxygen skal være > 60 % af værdien for mætning med luft i hele testperioden
One test-medium control series and also, if relevant, one control series containing the solvent or dispersant should be run in addition to the test series.
vandtemperaturen må ikke variere mere end ± 1,5 °C imellem testkamrene eller fra dag til dag på noget tidspunkt i testen, og den skal ligge i det temperaturområde, som gælder for den anvendte art (tillæg 2)
When used, the solvent or dispersant concentration should be the same as that used in the vessels containing the test chemical.
der skal foretages en analytisk måling af testkoncentrationerne
The appropriate number of replicates should be used (see paragraphs 23-24).
den generelle overlevelse af befrugtede æg og succesen efter klækning i kontrollerne og, hvor relevant, i opløsningsmiddelkontrollerne skal være større end eller lig med de i tillæg 2 fastsatte grænseværdier.
Generally in a well-run test, the coefficient of variation around the mean number of living offspring produced per parent animal in the control(s) should be ≤ 25 %, and this should be reported for test designs using individually held animals.
Hvis der konstateres en mindre afvigelse fra validitetskriterierne, bør konsekvenserne holdes op imod testdataenes pålidelighed, og disse overvejelser bør fremgå af rapporten.
Test medium renewal
Virkningerne på overlevelse, klækning eller vækst i opløsningsmiddelkontrollen, sammenlignet med den negative kontrol, bør rapporteres og drøftes i forbindelse med testdataenes pålidelighed.
The frequency of medium renewal will depend on the stability of the test chemical, but should be at least three times per week.
Testkamre
If, from preliminary stability tests (see paragraph 7), the test chemical concentration is not stable (i.e. outside the range 80 - 120 % of nominal or falling below 80 % of the measured initial concentration) over the maximum renewal period (i.e. 3 days), consideration should be given to more frequent medium renewal, or to the use of a flow-through test.
Enhver type glas, rustfrit stål eller andet inaktivt materiale kan anvendes.
When the medium is renewed in semi-static tests, a second series of test vessels are prepared and the parent animals transferred to them by, for example, a glass pipette of suitable diameter.
Eftersom silikone er kendt for en høj evne til absorbering af lipofile stoffer, bør brug af silikonerør i gennemstrømningsforsøg og brug af silikonelåg ved kontakt med vand minimeres, f.eks. ved brug af monoblokglasakvarier.
The volume of medium transferred with the Daphnia should be minimised.
Forsøgsbeholderne bør være store nok til at tillade tilstrækkelig vækst i kontrollen, opretholde koncentrationen af opløst oxygen (f.eks. vil et tankvolumen på 7 l være tilstrækkelig til små fiskearter) og opfylde de kriterier for belastningsgrad, der er anført i punkt 19.
The results of the observations made during the test should be recorded on data sheets (see examples in Appendixes 4 and 5).
Testkamrene bør anbringes randomiseret i testområdet.
If other measurements are required (see paragraph 44), additional observations may be required.
Et randomiseret blok-design, hvor samme behandling forefindes i hver blok, bør foretrækkes frem for et helt randomiseret design.
Offspring
Testkamrene bør skånes for unødvendige forstyrrelser.
The offspring produced by each parent animal should preferably be removed and counted daily from the appearance of the first brood to prevent them consuming food intended for the parent.