natgillin/translations-raw huggingface dataset:
kvaliteten af vandet i testglassene og krystallisationsskålene, dvs. pH, temperatur, opløst ilt, hårdhed og ammoniak
The food should be spread all over the soil surface and not just in one lump.
eventuel udskiftning af fordampet vand i testglassene
During transportation and during the testing period it should be avoided to knock or otherwise physically disturb the test containers, as this may increase the compaction of the soil, and hamper the interaction between the collembolans.
antal larver, der ikke udviklede sig til dansemyg, pr. glas for første og anden generation
A number of prerequisites should be fulfilled in advance before using alternative species:
procentvis/andel emergens pr. replikat og testkoncentration (samlet for han- og hunindivider) for første og anden generation
They should be unequivocally identified;
gennemsnitlig udviklingshastighed for fuldt udviklede og levende individer pr. replikat og behandlingshastighed (han- og hunindivider for sig og samlet) for første og anden generation
The rationale for the selection of the species should be given;
antal æggesnore afsat i krystallisationsskålene pr. avlsbur og pr. dag
It should be ensured that the reproductive biology is included in the testing phase so it will be a potential target during the exposure;
hver æggesnors karakteristika (størrelse, form og fertilitet)
The life-history should be known:
befrugtningsevne — samlet antal æggesnore pr. samlet antal hunner, der er tilført avlsburet
age at maturation, duration of egg development, and instars subject to exposure;
fertilitet — samlet antal fertile æggesnore pr. samlet antal hunner, der er tilført avlsburet
Optimal conditions for growth and reproduction should be provided by the test substrate and food supply;
estimater for toksicitetsendepunkter, f.eks. ECx-værdier (og tilhørende konfidensintervaller), NOEC og de statistiske metoder, der er anvendt til at beregningen heraf
Variability should be sufficiently low for precise and accurate toxicity estimation.
Kapitel C.28 i dette bilag, Toksicitetstest sediment/vand på chironomider med forurenet vand.
Extraction and counting of animals
Kapitel C.27 i dette bilag, Toksicitetstest sediment/vand på chironomider med forurenet sediment.
Two methods of extraction can be performed.
a Guidance to Application, OECD Series on Testing and Assessment No. 54, 146 s., ENV/JM/MONO(2006)18, OECD, Paris.
First method:
Formuleret sediment eller rekonstitueret, kunstigt eller syntetisk sediment en blanding af materialer, som bruges til at efterligne de fysiske bestanddele i naturligt sediment.
A controlled temperature gradient extractor based on principles by MacFadyen can be used (1).
Overliggende vand det vand, der tilsættes over sedimentet i testglasset.
The heat coming from a heating element at the top of the extraction box (regulated through a thermistor placed on the surface of the soil sample).
Interstitielt vand eller porevand det vand, der optager pladsen mellem sediment og jordpartikler.
The temperature in the cooled liquid surrounding the collecting vessel is regulated through a thermistor situated at the surface of the collection box (placed below the soil core).
Spiket vand eller forurenet vand det vand, som testkemikaliet er tilsat.
The thermistors are connected to a programmable controlling unit which raises the temperature according to a pre-programmed schedule.
Anbefalinger vedrørende dyrkning af Chironomus riparius
Animals are collected in the cooled collecting box (2 °C) with a bottom layer of plaster of Paris/charcoal.
Chironomus-larver kan holdes i krystallisationsskåle eller større beholdere.
Extraction is started at 25 °C and the temperature is increased automatically every 12 h by 5 °C and has a total duration of 48 hours.
Fint kvartssand spredes i et tyndt lag på ca. 5-10 mm på bunden af skålen.
After 12 h at 40 °C the extraction is finished.
Kieselgur (f.eks. Merck, Art 8117) er også et egnet substrat (et tyndere lag på få mm er nok).
Second method:
En passende mængde vand tilsættes, så der fås en dybde på flere cm.
After the experimental incubation period the number of juvenile Collembola present is assessed by flotation.
Vandet fyldes op efter behov for at erstatte tab som følge af fordampning og undgå udtørring.
For that purpose the test is performed in the vessels of approximately 250 ml volume.
Vandet kan udskiftes, hvis det er nødvendigt.
At the end of the test approx.
Forsigtig beluftning skal foretages.
200 ml of distilled water are added.
Larvebeholderne opbevares i et bur, der er egnet til formålet, og som forhindrer udviklede voksne i at forsvinde.
The soil is gently agitated with a fine paintbrush to allow Collembola to float to the water surface.
Buret skal være så stort, at udviklede voksne kan danne sværme.
A small amount, approx.
Ellers vil kopulation muligvis ikke forekomme (minimum er ca. 30 × 30 × 30 cm).
0,5 ml, of black Kentmere photographic dye may be added to the water to aid counting by increasing the contrast between the water and the white Collembola.
Burene opbevares ved rumtemperatur eller i rum med konstant temperatur ved 20 ± 2 °C med belysning, der består af 16 timers lys (intensitet ca. 1000 lux) og 8 timers mørke.
The dye is not toxic to Collembola.
Det er rapporteret, at en luftfugtighed på mindre end 60 % RH kan hæmme reproduktionen.
Counting:
Der kan anvendes egnet naturligt eller syntetisk vand.
Counts of numbers may be carried out by eye or under a light microscope using a grid placed over the floatation vessel or by photographing the surface of each vessel and later counting the Collembola on enlarged prints or projected slides.
Brøndvand, afkloret ledningsvand eller kunstige medier (f.eks. Elendt M4 eller M7, se nedenfor) anvendes ofte.
Counts may also be performed using digital image processing techniques (12).
Vandet beluftes inden brug.
All techniques should be validated.
Om nødvendigt kan vandet i beholderne udskiftes ved forsigtigt at hælde eller suge det brugte vand fra dyrkningsbeholderne uden at ødelægge larverørene.
Determination of the maximum WHC of the soil
Fodring af larver
The following method for determining the maximum water holding capacity (WHC) of the soil has been found to be appropriate.
Chironomus-larver fodres med fiskefoder i flager (Tetra Min®, Tetra Phyll® eller et tilsvarende mærke af særligt fiskefoder) med ca. 250 mg pr. beholder pr. dag.
It is described in Annex C of ISO DIS 11268-2 (Soil Quality — Effects of pollutants on earthworms (Eisenia fetida).
Foderet kan gives som tørt malet pulver eller som en suspension i vand:
Part 2: Determination of effects on reproduction).
1,0 g foder i flager tilsættes 20 ml fortyndet vand og blandes for at få en homogen blanding.
Cover the bottom of the tube with a wet piece of filter paper and then place it on a rack in a water bath.
Dette præparat kan tilsættes med ca. 5 ml pr. beholder pr. dag (rystes inden brug).
The water holding capacity (WHC) should be calculated as follows:
Ældre larver kan gives mere foder.
Soil Quality — Determination of pH.
Fodringen tilpasses vandkvaliteten.
C.40.
Hvis dyrkningsmediet bliver uklart, reduceres fodermængden.
SEDIMENT-WATER CHIRONOMID LIFE-CYCLE TOXICITY TEST USING SPIKED WATER OR SPIKED SEDIMENT
Tilsætningen af foder overvåges nøje.
This test method is equivalent to OECD Testing Guideline (TG) 233 (2010).
For lidt foder vil få larverne til at vandre til vandsøjlen, og for meget foder vil forårsage øget mikrobiel aktivitet og lavere iltkoncentrationer.
It is designed to assess the effects of life-long exposure of chemicals on the freshwater dipteran Chironomus sp., fully covering the 1st generation (P generation) and the early part of the 2nd generation (F1 generation).
Begge forhold kan medføre lavere vækstrater.
It is an extension of the existing test methods C.28 (1) or C.27 (15) using a spiked-water exposure scenario or a spiked sediment scenario, respectively.
Nogle grønalger (f.eks. Scenedesmus subspicatus og Chlorella vulgaris) kan også tilsættes ved etableringen af nye dyrkningsbeholdere.
It takes into account existing toxicity test protocols for Chironomus riparius and Chironomus dilutus (previously named C. tentans (2)) that have been developed in Europe and North America (3) (4) (5) (6) (7) (8) (9) and subsequently ring-tested (1) (7) (10) (11) (12).
Fodring af udviklede voksne
Other well documented chironomid species may also be used, e.g. Chironomus yoshimatsui (13) (14).
Nogle forskere har foreslået, at vat, der er opblødt i mættet rørsukkeropløsning, kan bruges som foder til udviklede voksne.
The complete exposure duration is ca.
Efter ca. 13-15 dage ved 20 ± 2 °C begynder voksne at udvikle sig fra larvebeholderne.
44 days for C. riparius and C. yoshimatsui, and –ca.
Hanner kan nemt identificeres ved deres fjerbuskede antenner og tynde krop.
100 days for C. dilutus.
Eventuelle ægmasser skal fjernes forsigtigt.
The water exposure scenario, spiking of the water column, is intended to simulate a pesticide spray drift event and covers the initial peak concentration in surface waters.
De overføres til en lille plade med en prøve af dyrkningsvandet.
Water spiking is also useful for other types of exposure (including chemical spills), but not for accumulation processes within the sediment lasting longer than the test period.
Ægmasser bruges til at starte et nyt dyrkningsglas (f.eks. 2-4 ægmasser pr. glas) eller til toksicitetstest.
In that case, and also when run-off is the main entry route of pesticides into water bodies, a spiked sediment design may be more appropriate.
Æglarver udklækkes efter 2-3 dage.
If other exposure scenarios are of interest, the test design may be readily adapted.
Etablering af nye dyrkningsglas
For example, if the distribution of the test chemical between the water phase and the sediment layer is not of interest and adsorption to the sediment has to be minimised, the use of surrogate artificial sediment (e.g. quartz sand) may be considered.
Når vækstbetingelser er etableret, kan et nyt larvedyrkningsglas etableres hver uge eller mindre hyppigt afhængigt af testkravene, således at de ældste glas kan fjernes, når voksne individer har udviklet sig.
Chemicals that require testing of sediment-dwelling organisms may persist in sediment over long periods.
Ved hjælp af dette system fås der er en regelmæssig forsyning af voksne med minimal håndtering.
Sediment-dwelling organisms may be exposed via a number of routes.
Fremstilling af testopløsninger M4 og M7
The relative importance of each exposure route, and the time taken for each to contribute to the overall toxic effect, is dependent on the physical-chemical properties of the chemical.
Elendt (1990) har beskrevet M4-mediet.
For strongly adsorbing chemicals or for chemicals covalently binding to sediment, ingestion of contaminated food may be a significant exposure route.
M7-mediet fremstilles på samme måde som M4-mediet bortset fra de stoffer, der er anført i tabel 1, for hvilke koncentrationerne er fire gange lavere i M7 end i M4.
In order not to underestimate the toxicity of highly lipophilic chemicals, the use of food added to the sediment before application of the test chemical may be considered (see paragraph 31).
I henhold til Elendt og Bias (1990) bør testopløsningen ikke fremstilles for de nævnte koncentrationer af NaSiO3 · 5H2O, NaNO3, KH2PO4 og K2HPO4, idet oplysningerne om fremstillingen af stamopløsninger ikke er fyldestgørende.
Therefore, it is possible to include all routes of exposure and all life stages.
Fremstilling af M7-mediet.
Measured endpoints are the total number of adults emerged (for both 1st and 2nd generations), development rate (for both 1st and 2nd generations), sex ratio of fully emerged and alive adults (for both 1st and 2nd generations), number of egg ropes per female (1st generation only) and fertility of the egg ropes (1st generation only).
Hver stamopløsning (I) fremstilles individuelt, og en kombineret stamopløsning (II) fremstilles ud fra disse stamopløsninger (I) (se tabel 1).
Formulated sediment is strongly recommended.
M7-mediet fremstilles ved at fortynde 50 ml af den kombinerede stamopløsning (II) og de mængder af hver stamopløsning med makronæringsstoffer, der er anført i tabel 2, med 1 liter deioniseret vand.
Formulated sediment has several advantages over natural sediments:
En vitaminopløsning fremstilles ved at tilsætte tre vitaminer til deioniseret vand som anført i tabel 3, og 0,1 ml af den kombinerede vitaminopløsning tilsættes det færdige M7-medium umiddelbart inden brug.
experimental variability is reduced because it forms a reproducible “standardised matrix” and the need to source uncontaminated clean sediment is eliminated;
Vitaminopløsningen opbevares i små aliquoter i frossen tilstand.
tests can be initiated at any time without encountering seasonal variability in the test sediment and there is no need to pre-treat the sediment to remove indigenous fauna;
Mediet beluftes og stabiliseres.
reduced cost compared to field collection of sufficient quantities required for routine testing;
Stamopløsninger af sporstoffer til M4- og M7-medierne
formulated sediment allows for comparisons of toxicity across studies and ranking chemicals accordingly (3).
Stamopløsninger (I)
First instar chironomid larvae are exposed to a concentration range of the test chemical in a sediment-water system.
Mængde (mg), der fortyndes med 1 l deioniseret vand
The test starts by placing first instar larvae (1st generation) into test beakers containing spiked sediment or alternately the test chemical is spiked into the water after addition of the larvae.
Fremstilling af den kombinerede stamopløsning (II):
Chironomid emergence, time to emergence and sex ratio of the fully emerged and alive midges are assessed.
Bland følgende mængder (ml) af stamopløsninger (I), og fortynd med 1 l deioniseret vand
Emerged adults are transferred to breeding cages, to facilitate swarming, mating and oviposition.
Endelige koncentrationer i testopløsninger (mg/l)
The number of egg ropes produced and their fertility are assessed.
MnCl2 · 4H2O [1]
From these egg ropes, first instar larvae of the 2nd generation are obtained.
SrCl2 · 6H2O [1]
These larvae are placed into freshly prepared test beakers (spiking procedure as for the 1st generation) to determine the viability of the 2nd generation through an assessment of their emergence, time to emergence and the sex ratio of the fully emerged and alive midges (a schematic presentation of the life-cycle test is provided in Appendix 5).
MoO4 · 2H2O [1]
All data are analysed either by a regression model to estimate the concentration that would cause X % reduction in the relevant endpoint, or by using hypothesis testing to determine a No Observed Effect Concentration (NOEC).
CuCl2 · 2H2O [1]
The latter requires a comparison of treatment responses with the appropriate control responses using statistical tests.
CaCl2 · 6H2O
It should be noted that in the spiked water scenario, in case of fast degrading chemicals, the later life stages of each generation (e.g. pupal phase) might be exposed to a considerably lower concentration level in the overlying water than the 1st instar larvae.
EDTA · 2H2O [1] [2]
If this is a concern, and a comparable exposure level for each life stage is needed, the following amendments of the test method might be considered:
FeSO4 · 7H2O [1] [2]
parallel runs with spiking at different life stages, or
Stamopløsninger med makro-næringsstoffer til M4- og M7-medierne
repeated spiking (or overlying water renewal) of the test system during both test phases (1st and 2nd generation), whereby the spiking (renewal) intervals should be adjusted to the fate characteristics of the test chemical.
Mængde, der fortyndes med 1 l deioniseret vand
Such amendments are only feasible in the spiked water scenario, but not in the sediment spiked scenario.
Mængde af stamopløsninger med makro-næringsstoffer, der tilsættes for at fremstille M4- og M7-medierne
The water solubility of the test chemical, its vapour pressure and log Kow, measured or calculated partitioning into sediment and stability in water and sediment should be known.
Endelige koncentrationer i testopløsninger M4 og M7
A reliable analytical method for the quantification of the test chemical in overlying water, pore water and sediment with known and reported accuracy and limit of detection should be available.
Vitaminopløsninger til M4- og M7-medierne
Useful information includes the structural formula and purity of the test chemical.
Alle tre vitaminopløsninger samles for at fremstille en vitaminopløsning.
Chemical fate of the test chemical (e.g. dissipation, abiotic and biotic degradation, etc.) is also useful.
Mængde af vitaminopløsning, der tilsættes for at fremstille M4- og M7-medierne
Further guidance for testing chemicals with physical-chemical properties that make them difficult to perform the test is provided in (16).
Thiaminhydrochlorid
Reference chemicals may be tested periodically as a means of assuring that the sensitivity of the laboratory population has not changed.
Biotin
As with daphnids it would be sufficient to perform a 48-h acute test (following 17).
Disse stoffer varierer i M4 og M7 som anført ovenfor.
However, until a validated acute guideline is available a chronic test according to Chapter C.28 of this Annex may be considered.
Disse opløsninger fremstilles hver for sig, hældes sammen og autoklaveres straks.
Examples of reference toxicants used successfully in ring-tests and validation studies are:
Det formulerede sediment skal have følgende sammensætning:
For the test to be valid the following conditions apply:
Sphagnum så nær pH 5,5-6,0 som muligt uden synlige planterester og findelt (partikelstørrelse ≤ 1 mm) og lufttørret
the mean emergence in the control treatment should be at least 70 % at the end of the exposure period for both generations (1) (7);
> 50 % af partiklerne skal være fra 50-200 μm
for C. riparius and C. yoshimatsui, 85 % of the total emerged adult midges from the control treatment in both generations should occur between 12 and 23 days after the insertion of the first instar larvae into the vessels;
CaCO3, pulveriseret, kemisk rent
for C. dilutus, a period of 20 to 65 days is acceptable;
0,05-0,1
the mean sex ratio of fully emerged and alive adults (as female or male fraction) in the control treatment of both generations should be at least 0,4, but not exceed 0,6; for each breeding cage the number of egg ropes in the controls of the 1st generation should be at least 0,6 per female added to the breeding cage;
Ledeevne ≤ 10 μS/cm
the fraction of fertile egg ropes in each breeding cage of the controls of the 1st generation should be at least 0,6; at the end of the exposure period for both generations, pH and the dissolved oxygen concentration should be measured in each vessel.
Suspensionen konditioneres i mindst to dage med forsigtig omrøring ved 20 °± 2 °C, for at stabilisere pH og opnå en stabil mikrobiel komponent. pH måles igen og skal være 6,0 ± 0,5.
The oxygen concentration should be at least 60 % of the air saturation value (ASV [9]), and the pH of overlying water should be between 6 and 9 in all test vessels;
Den færdige blandings pH måles igen og justeres om nødvendigt til 6,5 til 7,5 med CaCO3.
the water temperature should not differ by more than ± 1,0 °C.
Inden det formulerede sediment anvendes i en toksicitetstest på chironomider, bør det konditioneres i syv dage under de betingelser, der anvendes i den efterfølgende test.
Test vessels and breeding cages
OPBEVARING
The larvae are exposed in 600 ml glass beakers measuring ca.
De tørre bestanddele til fremstilling af syntetisk sediment opbevares tørt og køligt ved rumtemperatur.
8,5 cm in diameter (see Appendix 5).
Det formulerede (våde) sediment må ikke opbevares inden brug.
Other vessels are suitable, but they should guarantee a suitable depth of overlying water and sediment.
Det skal bruges umiddelbart efter konditioneringsperioden på de syv dage, som afslutter dets fremstilling.
The sediment surface should be sufficient to provide 2 to 3 cm2 per larvae.
Kemiske egenskaber for acceptabelt vand til fortynding
The ratio of the depth of the sediment layer to the depth of the overlying water should be ca.
BESTANDDEL
1:4.
KONCENTRATIONER
Breeding cages (minimum 30 cm in all three dimensions) with a gauze (mesh size ca.
Hårdhed som CaCO3
1 mm) on the top and one side of the cage as a minimum should be used (see Appendix 5).
Hvis der er formodning om en interaktion mellem hårdhedsioner og testkemikaliet, anvendes vand med lavere hårdhed (og dermed må Elendt-medium M4 ikke anvendes i denne situation).
In each cage a 2 l crystallising dish, containing test water and sediment, is placed for oviposition.
Vejledning i udførelse af test
Also for the crystallising dish, the ratio of the depth of the sediment layer to the depth of the overlying water should be around 1:4.
Eksempel på et bur
After egg ropes are collected from the crystallising dish they are placed into a 12-well microtiter plate (one rope per well containing at least 2,5 ml water from the spiked crystallising dish) after which the plates are covered with a lid to prevent significant evaporation.
A gaze øverst og på mindst den ene side af buret (maskestørrelse ca. 1 mm)
Other vessels suitable for keeping the egg ropes may also be used.
B åbning til anbringelse af udviklede voksne i buret og fjernelse af de lagte æggesnore fra krystallisationsskålene (vises ikke i denne grafik) C burstørrelse:
With the exception of the microtiter plates, all test vessels and other apparatus that will come into contact with the test system should be made entirely of glass or other chemically inert material (e.g. Polytetrafluoroethylene).
minimum 30 cm langt, 30 cm højt og 30 cm bredt
The species to be used in the test is preferably Chironomus riparius.
Eksempel på et testglas:
C. yoshimatsui may also be used.
A pasteurpipette til tilsætning af det overliggende vand med luft
C. dilutus is also suitable but more difficult to handle and requires a longer test period.
B glaslåg for at forhindre udviklede individer i at slippe ud
Details of culturing methods are given in Appendix 2 for C. riparius.
C vandoverflade
Information on culture conditions are also available for C. dilutus (5) and C. yoshimatsui (14).
D testglas (glasbæger, minimum 600 ml)
Identification of the species should be confirmed before testing but is not required prior to every test if the organisms come from an in-house culture.
E sedimentlag
Formulated sediment (also called reconstituted, artificial or synthetic sediment) should preferably be used.
Eksempel på en exhauster til at fange voksne individer (pilene viser luftstrømmens retning)
However, if natural sediment is used, it should be characterised (at least pH, organic carbon content, determination of other parameters such as C/N ratio and granulometry are also recommended) and should be free from any contamination and other organisms that may compete with, or consume chironomid larvae.
På indersiden er glasrørets åbning (A) dækket med noget vat og gaze (maskestørrelse ca. 1 mm2) for at hindre beskadigelse af individerne, når de suges ind i exhausteren
It is also recommended, before testing, that sediments are conditioned for seven days under test conditions.
C gennemsigtig beholder (plast eller glas, længde ca. 15 cm) til indfangede individer
The following formulated sediment, as described in (1), is recommended (1) (20) (21):
D prop af vulkaniseret gummi, perforeret med rør (E).
4-5 % (dry weight) peat:
Når individerne skal lukkes ind i buret, frigøres prop D fra beholder C
as close to pH 5,5 to 6,0 as possible;
E rør (plast eller glas, indvendig diameter ca. 8 mm) til at indsamle voksne individer fra glas
it is important to use peat in powder form, finely ground (particle size ≤ 1 mm) and only air dried;
Skematisk fremstilling af en livscyklustest:
20 % (dry weight) kaolin clay (kaolinite content preferably above 30 %);
A første generation — testglas indeholdende et sediment- og vandsystem, otte replikater, 20 æglarver pr. glas
75-76 % (dry weight) quartz sand (fine sand should predominate with more than 50 per cent of the particles between 50 and 200 μm);
B fire testglas til hvert bur, A og B
Deionised water is added to obtain moisture of the final mixture in the range of 30–50 %;