natgillin/translations-raw huggingface dataset:
komplet fortegnelse over levende afkom under testen efter hvert forældredyr (se eksempler på dataark i tillæg 4)
This k2 estimate can be used to estimate an assimilation efficiency according to equation A7.1, substituting the C0,d value in the equation with the measured time zero concentration (C0,m) in cases where C0,d is clearly estimated to be much higher than could have been achievable in the test.
antal døde forældredyr og dagen for hændelsen (se eksempler på dataark i tillæg 4)
If C0,m was not measureable, then the limit of detection in fish tissue should be used.
variationskoefficienten for kontrol af formeringsevne (baseret på det totale antal afkom pr. forældredyr, som er i live ved testens afslutning)
If, in some cases, this gives a value of α > 1, then the assimilation efficiency is assumed to 1 as a “worst case”.
det samlede antal levende afkom pr. forældredyr i hvert replikat bortset fra forældredyr, som kan være døde ved utilsigtet hændelse eller fejl i løbet af testen, plottet mod teststoffets koncentration
The maximum BMFK can then be estimated using Equation A7.4, and should be quoted as a “much less than” (<<) value.
hvor det er relevant, det samlede antal levende afkom pr. levende forældredyr i hvert replikat plottet mod testkemikaliets koncentration
For example, for a study carried out with a feeding rate of 3 % and a depuration half-life less than 3 days, and a “worst case” α of 1, the BMFK is likely to be below about 0,13.
hvor det er relevant, den laveste koncentration med observeret virkning (LOEC) med hensyn til formering, herunder en beskrivelse af de anvendte statistiske fremgangsmåder og en angivelse af, hvilken størrelse effekt der kan forventes påvist (der kan foretages en poweranalyse før forsøgets start) og koncentrationen uden observeret virkning (NOEC) med hensyn til formering; oplysninger om hvilken responsvariabel, der er anvendt til beregning af LOEC- og NOEC-værdien (enten som samlet antal levende afkom pr. moderorganisme, der ikke er død utilsigtet eller ved en fejl under forsøget, eller som det samlede antal levende afkom pr. overlevende moderorganisme); hvor det er relevant, rapporteres LOEC eller NOEC også for forældredyrenes dødelighed.
Given the purpose of this estimation and the fact that values will be conservative in nature, it is not necessary to correct them for growth dilution or fish and food lipid content.
hvor det er relevant, ECx for formering og konfidensintervaller (f.eks. 90 eller 95 %) og en graf over den model, der er anvendt til beregningen, hældningen på koncentration/respons-kurven og standardfejlen
In some regions it may only be possible to obtain fish food with a lipid concentration that falls far short of this upper limit.
andre observerede biologiske virkninger eller målinger.
In such cases studies should be run with the lower lipid concentration in the food as supplied, and the feeding rate adjusted appropriately to maintain fish health.
Der anføres enhver biologisk virkning, som er observeret eller målt (f.eks. forældredyrenes vækst), herunder relevante begrundelser
Diet lipids should not be artificially increased by the addition of excess oil.
en redegørelse for enhver afvigelse fra forsøgsmetoden.
Appendix 8
I forbindelse med denne forsøgsmetode anvendes følgende definitioner:
APPROACHES TO ESTIMATE TENTATIVE BCFS FROM DATA COLLECTED IN THE DIETARY EXPOSURE STUDY
Dødelighed ved en fejl
The dietary method is included in this test method for the bioaccumulation testing of substances that cannot in practice be tested using the aqueous exposure method.
Ikke kemikalierelateret dødelighed forårsaget af en utilsigtet hændelse (dvs. kendt årsag).
The aqueous exposure method gives a bioconcentration factor, whereas the dietary method leads directly to information on feeding biomagnification potential.
Et stof eller en blanding.
In many chemical safety regimes information on aquatic bioconcentration is required (for example in risk assessment and the Globally Harmonization System of Classification).
Den koncentration af testkemikaliet opløst i vand, som resulterer i en x procent reduktion i formeringsevne for Daphnia magna inden for en fastlagt eksponeringstid.
Hence there is a need to use the data generated in a dietary study to estimate a bioconcentration factor that is comparable to tests conducted according to the aqueous exposure method [1].
Utilsigtet dødelighed
This section explores approaches that may be followed to do this, while recognising the shortcomings that are inherent in the estimations.
Ikke kemikalierelateret dødelighed uden kendt årsag.
The dietary study measures depuration to give a depuration rate constant, k2.
Reel populationsforøgelse
If an uptake rate constant can be estimated with the available data for the situation where the fish had been exposed to the test substance via the water, then a kinetic BCF could be estimated.
Mål for vækst, der integrerer formeringsevne og aldersbetinget dødelighed (1)(2)(3).
The estimation of an uptake rate constant for water exposure of a test substance is reliant on many assumptions, all of which will contribute to the estimate's uncertainty.
I en ligevægtspopulation vil den være lig nul.
Furthermore, this approach to estimating a BCF assumes that the overall rate of depuration (including contributory factors like distribution in the body and individual depuration processes) is independent of the exposure technique used to produce a test substance body burden.
For populationer i vækst vil den være positiv, og for populationer i nedgang vil den være negativ.
The main assumptions inherent in the estimation approach can be summarised as follows.
Det er klart, at den sidste situation er uholdbar og vil ende med populationens udslettelse.
Depuration following dietary uptake is the same as depuration following aqueous exposure for a given substance
Bestemmelsesgrænse
uptake can be correlated with fish weight alone
Den laveste koncentration, der kan måles kvantitativt.
uptake can be correlated with the substance's octanol-water partition coefficient alone
Laveste koncentration med observeret effekt (LOEC)
uptake can be correlated with a combination of fish weight and the substance's octanol-water partition coefficient
Den laveste kemikaliekoncentration, der i testen har vist sig at have en statistisk signifikant indflydelse (p < 0,05) på formering og forældredyrs dødelighed inden for den fastlagte eksponeringstid, sammenlignet med kontrollen.
factors that can affect uptake in an aqueous exposure study in practice such as substance bioavailability, adsorption to apparatus, molecular size etc. have little effect
Imidlertid skal alle undersøgte koncentrationer, der ligger over LOEC, have haft en skadelig virkning, som er større end eller lig med den, som er observeret ved LOEC.
and, crucially:
Er disse to betingelser ikke opfyldt, bør der fyldestgørende redegøres for, hvordan den pågældende LOEC (og dermed NOEC) er valgt.
The database (“training set”) used to develop the uptake estimation method is representative of the substance under consideration
Et dyr registreres som dødt, når det ikke bevæger sig, dvs. når det ikke er i stand til at svømme, eller hvis der ikke ses bevægelser af vedhæng eller postabdomen inden for 15 sekunder efter forsigtig omrystning af testbeholderen.
Several publications in the open literature have derived equations relating uptake from water in fish via the gills to a substance's octanol-water partition coefficient, fish weight (1) (2) (3) (4), volume and/or lipid content, membrane permeation/diffusion (5) (6), fish ventilation volume (7) and by a fugacity/mass balance approach (8) (9) (10).
(Benyttes en anden definition, bør dette angives tillige med den tilhørende henvisning).
A detailed appraisal of such methods in this context is given in Crookes & Brooke (11).
Koncentration uden observeret effekt (NOEC)
A publication by Barber (12) focussed on modelling bioaccumulation through dietary uptake is also useful in this context as it includes contributions from gill uptake rate models.
Den testkoncentration umiddelbart under LOEC, som ved sammenligning med kontrollen ikke har nogen statistisk signifikant virkning (p < 0,05) inden for en fastlagt eksponeringstid.
A section of the background document to the 2004 dietary protocol (13) was also devoted to this aspect.
Unge dafnier, der er født af forældredyr under testen.
Most of these models seem to have been derived using limited databases.
Forældredyr
For models where details of the database used to build the model are available, it appears that the types of substances used are often of a similar structure or class (in terms of functionality, e.g. organochlorines).
Hundafnier, som er til stede ved testens begyndelse, og hvis formeringsevne er genstand for undersøgelse.
This adds to the uncertainty in using a model to predict an uptake rate constant for a different type of substance, in addition to test-specific considerations like species, temperature, etc.
Formeringsevne
A review of available techniques (11) highlighted that no one method is “more correct” than the others.
Antal levende afkom pr. forældredyr inden for testperioden.
Therefore, a clear justification should be given for the model used.
Et stof eller en blanding, som testes ved hjælp af denne forsøgsmetode.
Where several methods are available for which the use can be justified, it may be prudent to present several estimates of k1 (and so BCF) or a range of k1 values (and BCF) according to several uptake estimation methods.
LITTERATUR
However, given the differences in model types and datasets used to develop them, taking a mean value from estimates derived in different ways would not be appropriate.
KOMPLET FREMSTILLING AF MEDIERNE ELENDT M7 OG M4
Some researchers have postulated that BCF estimates of this sort require a bioavailability correction to account for a substance's adsorption to dissolved organic carbon (DOC) under aqueous exposure conditions, to bring the estimate in line with results from aqueous exposure studies (e.g. (13) (14)).
Akklimatisering til medierne Elendt M7 og M4
Howeverl this correction may not be appropriate given the low levels of DOC required in an aqueous exposure study for a ‘worst case’ estimate (i.e. ratio of bioavailable substance to substance as measured in solution).
Nogle laboratorier har haft problemer med at overflytte dafnier direkte til M4- (1) og M7-mediet.
For highly hydrophobic substances uptake at the gill may become limited by the rate of passive diffusion near the gill surface; in this case it is possible that the correction may be accounting for this effect rather than what it was designed for.
Dog har der været en vis succes med gradvis akklimatisering, dvs. flytning fra eget medium til 30 % Elendt efterfulgt af 60 % Elendt og derefter 100 % Elendt.
It is advised to focus on methods that require inputs for which data will be readily available for substances tested according to the dietary study described here (i.e. log KOW, fish weight).
En akklimatiseringsperiode på helt op til en måned kan være nødvendig.
Other methods that require more complex inputs may be applied, but may need additional measurements in the test or detailed knowledge on the test substance or fish species that may not be widely available.
Fremstilling
In addition, choice of model may be influenced by the level of validation and applicability domain (see (11) for a review and comparison of different methods).
Sporstoffer
It should be borne in mind that the resulting k1 estimate, and estimated BCF, are uncertain and may need to be treated in a weight-of-evidence approach along with the derived BMF and substance parameters (e.g. molecular size) for an overall picture of a substance's bioaccumulation potential.
Først fremstilles separate stamopløsninger (I) af hvert mikronæringsstof i vand af passende renhed, f.eks. deioniseret eller destilleret vand eller vand behandlet ved omvendt osmose.
Interpretation and use of these parameters may depend on the regulatory framework.
Af disse forskellige stamopløsninger fremstilles I) en enkelt stamopløsning II) indeholdende alle mikronæringsstoffer (kombineret opløsning), dvs.:
Sijm D.T.H.M., Verberne M.E., de Jonge W.J., Pärt P. and Opperhuizen A. (1995), Allometry in the uptake of hydrophobic chemicals determined in vivo and in isolated perfused gills.
Stamopløsning(er) I
In the wild the route leading to greatest exposure in aqueous environments is likely to be through ingestion for very hydrophobic substances and so an estimated BCF is not strictly representative of such a substance's bioaccumulation potential.
(et enkelt stof)
In Part C, Chapter C.20 is replaced by the following:
Mængde tilsat til vand
Daphnia magna Reproduction Test
Koncentration (i forhold til M4-medium)
This test method is equivalent to OECD test guideline (TG) 211 (2012).
Til fremstilling af den kombinerede stamopløsning II tilsættes følgende mængde stamopløsning I til vand
OECD test guidelines are periodically reviewed in the light of scientific progress.
gammel
The reproduction test guideline 211 originates from test guideline 202, Part II, Daphnia sp. reproduction test (1984).
De to opløsninger af Na2EDTA og FeSO4 fremstilles hver for sig, hældes sammen og autoklaveres straks.
It had generally been acknowledged that data from tests performed according to that TG 202 could be variable.
Derved fås:
This led to considerable effort being devoted to the identification of the reasons for this variability with the aim of producing a better test method.
Fe-EDTA-opløsning
Test guideline 211 is based on the outcome of these research activities, ring-tests and validation studies performed in 1992 (1), 1994 (2) and 2008 (3).
M4- og M7-medium
The main differences between the initial version (TG 202, 1984), and second version (TG 211, 1998) of the reproduction test guideline are:
M4- og M7-medierne fremstilles af stamopløsning II, makronæringsstoffer og vitaminer på følgende måde:
the recommended species to be used is Daphnia magna;
Mængde stamopløsning tilsat til fremstilling af mediet
the test duration is 21 days;
Stamopløsning II
for semi-static tests, the number of animals to be used at each test concentration has been reduced from at least 40, preferably divided into four groups of 10 animals, to at least 10 animals held individually (although different designs can be used for flow-through tests);
(kombinerede sporstoffer)
more specific recommendations have been made with regard to test medium and feeding conditions.
Stamopløsninger af makronæringsstoffer (enkelt stof)
the main differences between the second version of the reproduction test guideline (TG 211, 1998) and this version are:
Kombineret vitamin-stamopløsning
appendix 7 has been added to describe procedures for the identification of neonate sex if required.
Den kombinerede vitamin-stamopløsning fremstilles ved tilsætning af de tre vitaminer til 1 liter vand som vist nedenfor:
In line with previous versions of this test method sex ratio is an optional endpoint;
Thiaminhydrochlorid
the response variable number of living offspring produced per surviving parental animal has been supplemented with an additional response variable for Daphnia reproduction, i.e. the total number of living offspring produced at the end of the test per parent daphnia at the start of the test excluding from the analysis parental accidental and/or inadvertent mortality.
Cyanocobalamin (B12)
The purpose of the added response variable is to align this response variable with other reproduction test methods on invertebrates.
Biotin
Furthermore, in relation to this response variable, it is possible, in this test method, to remove a source of error, namely the effect of inadvertent and/or accidental parental mortality, should that occur during the exposure period.
Den kombinerede vitamin-stamopløsning opbevares nedfrosset i små alikvoter.
additional statistical guidance for test design and for treatment of results has been included both for ECx (e.g. EC10 or EC50) and for NOEC/LOEC approach.
Vitaminerne tilsættes mediet kort før brug.
a limit test is introduced.
For at undgå udfældning af salte, når det fuldstændige medium fremstilles, tilsættes de enkelte opløsninger til 500-800 ml deioniseret vand, hvorefter der ihældes op til 1 liter.
Definitions used are given in Appendix 1.
Den første publicerede beskrivelse af M4-mediet kan findes i Elendt, B.P.
The primary objective of the test is to assess the effect of chemicals on the reproductive output of Daphnia magna.
ANALYSE AF DET TOTALE ORGANISKE KULSTOF (TOC) OG FREMSTILLING AF NOMOGRAM TIL TOC-INDHOLDET I ALGEFODERET
To this end, young female Daphnia (the parent animals), aged less than 24 hours at the start of the test, are exposed to the test chemical added to water at a range of concentrations.
Det erkendes, at kulstofindholdet i algefoderet almindeligvis ikke vil blive målt direkte, men ud fra korrelationer (dvs. nomogrammer med erstatningsmålinger, såsom antallet af algeceller eller lysabsorbans).
The test duration is 21 days.
TOC bør måles ved oxidering ved høje temperaturer og ikke ved UV- eller persulfatmetoder.
At the end of the test, the total number of living offspring produced is assessed.
(Uddybende oplysninger findes i: The Instrumental Determination of Total Organic Carbon, Total Oxygen Demand and Related Determinands 1979, HMSO 1980; 49 High Holborn, London WC1V 6HB).
Reproductive output of the parent animals can be expressed in other ways (e.g. number of living offspring produced per animal per day from the first day offspring were observed) but these should be reported in addition to the total number of living offspring produced at the end of the test.
Med henblik på fremstilling af nomogrammer separeres algerne fra dyrkningsmediet ved centrifugering med efterfølgende resuspension i destilleret vand.
Because of the particular design of the semi-static test compared to other invertebrate reproduction test methods, it is also possible to count the number of living offspring produced by each individual parent animal.
Erstatningsparameteren og TOC-koncentrationen måles i hver prøve som en tredobbelt bestemmelse.
This enables that, contrary to other invertebrate reproduction test methods, if the parent animal dies accidentally and/or inadvertently during the test period, its offspring production can be excluded from data assessment.
Blindværdier måles i destilleret vand, og TOC-koncentrationen trækkes fra TOC-værdien i algeprøven.
Hence, if parental mortality occurs in exposed replicates, it should be considered whether or not the mortality follows a concentration-response pattern, e.g. if there is a significant regression of the response versus concentration of the test chemical with a positive slope (a statistical test like the Cochran-Armitage trend test may be used for this).
Nomogrammerne skal være lineære i hele området over kulkoncentrationer.
If the mortality does not follow a concentration-response pattern, then those replicates with parental mortality should be excluded from the analysis of the test result.
Eksempler ses nedenfor:
If the mortality follows a concentration-response pattern, the parental mortality should be assigned as an effect of the test chemical and the replicates should not be excluded from the analysis.
Disse bør ikke bruges til konvertering; det er essentielt, at laboratorierne selv fremstiller deres egne nomogrammer.
If the parent animal dies during the test i.e. accidentally from mishandling or accident, or inadvertently due to unexplained incident not related to the effect of the test chemical or turns out to be male, then the replicate is excluded from the analysis (see more in paragraph 51).
Regression af mg/l tørvægt på mg C/1.
The toxic effect of the test chemical on reproductive output is expressed as ECx by fitting the data to an appropriate model by non-linear regression to estimate the concentration that would cause x % reduction in reproductive output, respectively, or alternatively as the NOEC/LOEC value (4).
Data fra koncentrerede suspensioner af semikontinuerligt dyrkede celler, der genopslæmmes i destilleret vand.
The test concentrations should preferably bracket the lowest of the used effect concentrations (e.g. EC10) which means that this value is calculated by interpolation and not extrapolation.
x-akse: mg C/1 af koncentreret algefoder
The survival of the parent animals and time to production of first brood should also be reported.
y-akse: mg/1 tørvægt af koncentreret algefoder
Other chemical-related effects on parameters such as growth (e.g. length), and possibly intrinsic rate of population increase, can also be examined (see paragraph 44).
Korrektionskoefficient -0,980
INFORMATION ON THE TEST CHEMICAL
Regression af celleantal på mg C/1.
Results of an acute toxicity test (see chapter C.2 of this Annex: Daphnia sp. acute immobilisation test) performed with Daphnia magna may be useful in selecting an appropriate range of test concentrations in the reproduction tests.
y-akse: antal celler/1 af koncentreret algefoder
The water solubility and the vapour pressure of the test chemical should be known and a reliable analytical method for the quantification of the chemical in the test solutions with reported recovery efficiency and limit of determination should be available.
korrektionskoefficient -0,926
Information on the test chemical which may be useful in establishing the test conditions includes the structural formula, purity of the chemical, stability in light, stability under the conditions of the test, pKa, Pow and results of a test for ready biodegradability (see chapters C.4 (determination of ‘ready’ biodegradability), and C.29 (ready biodegradability — CO2 in sealed vessels) of this Annex).
Regression af absorbans på mg C/1 (1 cm stilængde).
For a test to be valid, the following performance criteria should be met in the control(s):
y-akse: Absorbansen ved 440 nm af en 1/10-fortynding af koncentreret algefoder
the mortality of the parent animals (female Daphnia) does not exceed 20 % at the end of the test;
Korrektionskoefficient – 0,998
the mean number of living offspring produced per parent animal surviving at the end of the test is ≥ 60.
EKSEMPEL PÅ DATAARK TIL NOTATER OM UDSKIFTNING AF MEDIUM, FYSISK/KEMISK MONITORERING, FODRING, FORMERING OG DØDELIGHED BLANDT FORÆLDREDYR
Note: The same validity criterion (20 %) can be used for accidental and inadvertent parental mortality for the controls as well as for each of the test concentrations.
Forsøg nr.:
Test vessels and other apparatus, which will come into contact with the test solutions, should be made entirely of glass or other chemically inert material.
Startdato:
The test vessels will normally be glass beakers.
Klon:
In addition, some or all of the following equipment will be required:
Type foder:
oxygen meter (with microelectrode or other suitable equipment for measuring dissolved oxygen in low volume samples);
Testkemikalie:
adequate apparatus for temperature control;
Nominel konc.:
pH-meter;
Dag
equipment for the determination of the hardness of water;
Udskiftning af testmedium (sæt kryds)
equipment for the determination of the total organic carbon concentration (TOC) of water or equipment for the determination of the chemical oxygen demand (COD);
gammel
adequate apparatus for the control of the lighting regime and measurement of light intensity.
O2 (mg/l) [1] ny
Test Organism
Foder (sæt kryds)
The species to be used in the test is Daphnia magna Straus [57].Preferably, the clone should have been identified by genotyping.
Antal levende afkom [2]
Research (1) has shown that the reproductive performance of Clone A (which originated from IRCHA in France) (5) consistently meets the validity criterion of a mean of ≥ 60 living offspring per parent animal surviving when cultured under the conditions described in this test method.
I alt
However, other clones are acceptable provided that the Daphnia culture is shown to meet the validity criteria for the test.
Beholder 1
At the start of the test, the animals should be less than 24 hours old and should not be first brood progeny.
Kumulativ dødelighed for forældredyr [3]
They should be derived from a healthy stock (i.e. showing no signs of stress such as high mortality, presence of males and ephippia, delay in the production of the first brood, discoloured animals, etc.).