natgillin/translations-raw huggingface dataset:
Primære endepunkter / Bestemmelsesdage
Analytical Chemistry
D 7 og 21
Prior to conducting a study, the stability of the test chemical should be evaluated using existing information on its solubility, degradability and volatility.
Histologisk undersøgelse af thyreoidea
Test solutions from each replicate tank at each concentration should be sampled for analytical chemistry analyses at test initiation (day 0), and weekly during the test for a minimum of four samples.
Fortyndingsvand / Laboratoriekontrol
It is also recommended that each test concentration be analysed during system preparation, prior to test initiation, to verify system performance.
Afkloret ledningsvand (trækulsfiltreret) eller tilsvarende laboratoriekilde
In addition, it is recommended that stock solutions be analysed when they are changed, especially if the volume of the stock solution does not provide adequate amounts of chemical to span the duration of routine sampling periods.
Larvetæthed
In the case of chemicals which cannot be detected at some or all of the concentrations used in a test, stock solutions should be measured and system flow rates recorded in order to calculate nominal concentrations.
20 larver / testbeholder (5 / l)
Chemical Delivery
Testopløsning / Testbeholder
The method used to introduce the test chemical to the system can vary depending on its physicochemical properties.
4-10 l (minimum 10-15 cm vand) / Testbeholder af glas eller rustfrit stål (f.eks. 22,5 cm × 14 cm × 16,5 cm)
Water soluble chemicals can be dissolved in aliquots of test water at a concentration which allows delivery at the target test concentration in a flow-through system.
Gengivelse
Chemicals which are liquid at room temperature and sparingly soluble in water can be introduced using liquid:liquid saturator methods.
4 testbeholdere til replikat / testkoncentration og kontrolprøve
Chemicals which are solid at room temperature and are sparingly soluble in water can be introduced using glass wool column saturators (7).
Acceptabel dødelighed i kontrollerne
The preference is to use a carrier-free test system, however different test chemicals will possess varied physicochemical properties that will likely require different approaches for preparation of chemical exposure water.
≤ 10 % pr. replikattestbeholder
It is preferred that effort be made to avoid solvents or carriers because:
Fiksering af skjoldbruskkirtel
certain solvents themselves may result in toxicity and/or undesirable or unexpected endocrinological responses, ii) testing chemicals above their water solubility (as can frequently occur through the use of solvents) can result in inaccurate determinations of effective concentrations, and iii) the use of solvents in longer-term tests can result in a significant degree of “biofilming” associated with microbial activity.
Fikseret antal
For difficult to test chemicals, a solvent may be employed as a last resort, and the OECD Guidance Document on aquatic toxicity testing of difficult substances and mixtures should be consulted (4) to determine the best method.
Alle haletudser (5/replikat vurderes først)
Solvents which have been found to be effective for aquatic toxicity testing include acetone, ethanol, methanol, dimethyl formamide and triethylene glycol.
Område
In case a solvent carrier is used, solvent concentrations should be below the chronic No Observed Effect Concentration (NOEC);
Hoved eller hele kroppen
the OECD Guidance Document recommends a maximum of 100 μl/l;
Fikseringsmiddel
a recent review recommends that solvent concentrations as low as 20 μl/l of dilution water be used (12).
Davidson's fikseringsmiddel
If solvent carriers are used, appropriate solvent controls should be evaluated in addition to non-solvent controls (clean water).
Foder
If it is not possible to administer a chemical via the water, either because of physicochemical characteristics (low solubility) or limited chemical availability, introducing it via the diet may be considered.
Sera Micron® eller tilsvarende
Preliminary work has been conducted on dietary exposures;
Mængde / Frekvens
however, this route of exposure is not commonly used.
Se tabel 1 for fodringhyppighed med Sera Micron®
The choice of method should be documented and analytically verified.
Belysning
Establishing the High Test Concentration
Lysperiode
For the purposes of this test, the high test concentration should be set by the solubility limit of the test chemical;
12 timers lys:
the maximum tolerated concentration (MTC) for acutely toxic chemicals;
12 timers mørke
or 100 mg/l, whichever is lowest.
Intensitet
The MTC is defined as the highest test concentration of the chemical which results in less than 10 % acute mortality.
600-2000 lux (målt ved vandoverfladen)
Using this approach assumes that there are existing empirical acute mortality data from which the MTC can be estimated.
Vandtemperatur
Although the use of regression models may be the most technically sound approach to estimating the MTC, a useful approximation of the MTC can be derived from existing acute data by using 1/3 of the acute LC50 value.
6,5-8,5
However, acute toxicity data may be lacking for the species on test.
Koncentration af opløst ilt (DO)
If species specific acute toxicity data are not available, then a 96-hour LC50 test can be completed with tadpoles that are representative (i.e., same stage) of those on test in the AMA.
> 3,5 mg/l (> 40 % luftmætning)
Optionally, if data from other aquatic species are available (e.g. LC50 studies in fish or other amphibian species), then professional judgment may be used to estimate a likely MTC based on inter-species extrapolation.
Tidsplan for kemiske analyseprøver
Alternatively, if the chemical is not acutely toxic and is soluble above 100 mg/l, then 100 mg/l should be considered the highest test concentration (HTC), as this concentration is typically considered “practically non-toxic.”
En gang om ugen (4 prøver pr. test)
Although not the recommended procedure, static renewal methods may be used where flow-through methods are inadequate to achieve the MTC.
Indberetningsskemaer til rådata og oversigtsdata
If static renewal methods are used, then the stability of the test chemical concentration should be documented and remain within the performance criteria limits.
Generelle oplysninger om testkemikalie
Twenty-four hour renewal periods are recommended.
Kemisk information
Renewal periods exceeding 72 hours are not acceptable.
Indfør testkemikalie, koncentrationsenheder og behandlinger
Additionally, water quality parameters (e.g. DO, temperature, pH, etc.) should be measured at the end of each renewal period, immediately prior to renewal.
Koncentrationsenheder:
Test Concentration Range
Behandling 1
There is a required minimum of three test concentrations and a clean water control (and vehicle control if necessary).
Behandling 2
The minimum test concentration differential between the highest and lowest should be about one order of magnitude.
Behandling 3
The maximum dose separation is 0,1 and the minimum is 0,33.
Behandling 4
Test Initiation and Conduct
Dato (dag 0):
The exposure should be initiated when a sufficient number of tadpoles in the pre-exposure stock population have reached developmental stage 51, according to Nieuwkoop and Faber (8), and which are less than or equal to 17 days of age post fertilisation.
Indfør dato (mm/dd/åå)
For selection of test animals, healthy and normal looking tadpoles of the stock population should be pooled in a single vessel containing an appropriate volume of dilution water.
Dato (dag 7):
For developmental stage determination, tadpoles should be individually removed from the pooling tank using a small net or strainer and transferred to a transparent measurement chamber (e.g. 100 mm Petri dish) containing dilution water.
Dato (dag 21):
For stage determination, it is preferred not to use anaesthesia, however one may individually anaesthetise the tadpoles using 100 mg/l tricaine methanesulfonate (e.g. MS-222), appropriately buffered with sodium bicarbonate (pH 7,0), before handling.
Skemaer til indsamling af rådata for dag 7 og dag 21
If used, methodology for appropriately using e.g. MS-222 for anaesthesia should be obtained from experienced laboratories and reported with the test results.
DAG X
Animals should be carefully handled during this transfer in order to minimise handling stress and to avoid any injury.
DATO 00/00/00
The developmental stage of the animals is determined using a binocular dissection microscope.
Behandlings-nummer
To reduce the ultimate variability in developmental stage, it is important that this staging be conducted as accurately as possible.
Replikat-nummer
According to Nieuwkoop and Faber (8), the primary developmental landmark for selecting stage 51 organisms is hind limb morphology.
Individuelt nummer
The morphological characteristics of the hind limbs should be examined under the microscope.
Individuel indikator
While the complete Nieuwkoop and Faber (8) guide should be consulted for comprehensive information on staging tadpoles, one can reliably determine stage using prominent morphological landmarks.
SVL-længde (mm)
The following table can be used to simplify and standardise the staging process throughout the study by identifying those prominent morphological landmarks associated with different stages, assuming that development is normal.
Baglemslængde (mm)
Prominent morphological staging landmarks based on Neuwkoop and Faber guidance
Hele organismens vægt (våd) (mg)
Prominent Morphological Landmarks
Beregnede oversigter for endepunktsdata fra dag 7 og dag 21
Developmental Stage
Celleberegninger er knyttet til dataindførsler i tabel 2
Hindlimb
Vægt (mg)
Forelimb
GNSN
Craniofacial structure
Daglige data om dødelighed
Olfactory nerve morphology
Celleberegninger er knyttet til dataindførsler i tabel 1.
Tail length
Testdag
For test initiation, all tadpoles should be at stage 51.
Optælling af replikater
The most prominent morphological staging landmark for that stage is hind limb morphology, which is demonstrated in Figure 1.
Optælling af behandlinger
Hind limb morphology of a stage 51 X. laevis tadpole
Tabel 5
In addition to the developmental stage selection, an optional size selection of the experimental animals may be used.
Vandkvalitetskriterier
For this purpose, the whole body length (not SVL) should be measured at day 0 for a sub-sample of approximately 20 NF stage 51 tadpoles.
Eksponeringssystem (gennemstrømning/statisk udskiftning):
After calculation of the mean whole body length for this group of animals, minimum and maximum limits for the whole body length of experimental animals can be set by allowing a range of the mean value ± 3 mm (mean values of whole body length range between 24,0 and 28,1 mm for stage 51 tadpoles).
Lysintensitet:
However, developmental staging is the primary parameter in determining the readiness of each test animal.
Lys-/mørkecyklus:
Tadpoles exhibiting grossly visible malformations or injuries should be excluded from the assay.
Foder:
Tadpoles that meet the stage criteria described above are held in a tank of clean culture water until the staging process is completed.
Fodermængde:
Once the staging is completed, the larvae are randomly distributed to exposure treatment tanks until each tank contains 20 larvae.
Vandets pH:
Each treatment tank is then inspected for animals with abnormal appearance (e.g., injuries, abnormal swimming behaviour, etc.).
Iodkoncentration i testvandet:
Overtly unhealthy looking tadpoles should be removed from the treatment tanks and replaced with larvae newly selected from the pooling tank.
Tabel 6
For more in-depth information on test termination procedures and processing of tadpoles, refer to the OECD Guidance Document on Amphibian Thyroid Histology (9).
Tabel 7
The random procedure used should give each organism on test equal probability of being selected.
Dato:
Tadpoles not selected are returned to the tank of origin and the selected tadpoles are humanely euthanised in 150 to 200 mg/l e.g. MS-222, appropriately buffered with sodium bicarbonate to achieve pH 7,0.
Kemikalie:
The euthanised tadpoles are rinsed in water and blotted dry, followed by body weight determination to the nearest milligram.
Patolog:
Hind limb length, snout to vent length, and developmental stage (using a binocular dissection microscope) are determined for each tadpole.
Forstørret skjoldbruskkirtel (hypertrofi)
Day 21 Measurements (Test Termination)
Formindsket skjoldbruskkirtel (atrofi)
At test termination (day 21), the remaining tadpoles are removed from the test tanks and humanely euthanised in 150 to 200 mg/l e.g. MS-222, appropriately buffered with sodium bicarbonate, as above.
Follikelcellehypertrofi
Tadpoles are rinsed in water and blotted dry, followed by body weight determination to the nearest milligram.
Follikelcellehyperplasi
Developmental stage, SVL, and hind limb lengths are measured for each tadpole.
Kontroldyr ID — replikat 1
All larvae are placed in Davidson's fixative for 48 to 72 hours either as whole body samples or as trimmed head tissue samples containing the lower jaw for histological assessments.
Dosis dyr ID — replikat 1
For histopathology, a total of five tadpoles should be sampled from each replicate tank.
Kontroldyr ID — replikat 2
Since follicular cell height is stage dependent (10), the most appropriate sampling approach for histological analyses is to use stage-matched individuals, whenever possible.
Dosis dyr ID — replikat 2
In order to select stage-matched individuals, all larvae should first be staged prior to selection and subsequent processing for data collection and preservation.
i alt:
This is necessary because normal divergence in development will result in differential stage distributions within each replicate tank.
Tabel 8
Animals selected for histopathology (n = 5 from each replicate) should be matched to the median stage of the controls (pooled replicates) whenever possible.
Supplerende histopatologiske kriterier
If there are replicate tanks with more than five larvae at the appropriate stage, then five larvae are randomly selected.
Tabel 9
That is, if the chemical treatment is associated with a retardation of development, then additional larvae should be sampled from the next lower stage.
Forklarende beskrivelser for histopatologiske fund
In turn, if the chemical treatment is associated with an acceleration of development, then additional larvae should be sampled from the next upper stage.
Forklarende beskrivelse
In cases of severe alterations of tadpole development due to treatment with a test chemical, there might be no overlap of the stage distribution in the chemical treatments with the calculated control median developmental stage.
Tabel 10
In only these cases, the selection process should be modified by using a stage different from the control median stage to achieve a stage-matched sampling of larvae for thyroid histopathology.
Oversigt over skabelon til indberetningsskema for dag x (7 eller 21) i AMA
Furthermore, if stages are indeterminate (i.e., asynchrony), then 5 tadpoles from each replicate should be randomly chosen for histological analysis.
Kontrol
The rationale underlying sampling of any larvae that are not at a stage equivalent to the control median developmental stage should be reported.
Dosis 1
Determination of Biological Endpoints
Dosis 2
During the 21 day exposure phase, measurement of primary endpoints is performed on days 7 and 21, however daily observation of test animals is necessary.
Dosis 3
Table 3 provides an overview of the measurement endpoints and the corresponding observation time points.
Endepunkt
More detailed information for technical procedures for measurement of apical endpoints and histological assessments is available in the OECD guidance documents (9).
Replikat
Observation time points for primary endpoints in the AMA
Middel
Apical Endpoints
p-værdi
Daily
Middel:
Mortality
Vådvægt
Hind Limb Length
Tabel 11
Snout-Vent Length
Oversigt over skabelon til indberetningsskema for dag x (7 eller 21) udviklingsstadiedata for AMA
Wet Body Weight
Alternativ analyse af vægt og længde i tilfælde af, at sen stadieudvikling gælder for over 20 % af haletudserne i en eller flere koncentration(er)
Thyroid Gland Histology
Såfremt et relativt stort antal haletudser udviser en udvikling over stadium 60 (≥ 20 %) i en eller flere nominelle koncentration(er), bør der foretages en tofaktor-ANOVA med en indlejret variansstruktur på alle haletudser for at vurdere vækstvirkninger som følge af kemisk behandling, samtidig med at der tages hensyn til, hvordan udvikling i et sent stadium påvirker væksten.
Developmental stage, hind limb length, SVL and wet weight are the apical endpoints of the AMA, and each is briefly discussed below.
Det foreslås at bruge alle data, men tage hensyn til virkningen af udvikling i et sent stadium.
Further technical information for collecting these data is available in the guidance documents referenced including procedures for computer-assisted analysis which are recommended for use.
Dette kan gøres med en tofaktor-ANOVA med en indlejret variansstruktur.
The developmental stage of X. laevis tadpoles is determined using the staging criteria of Nieuwkoop and Faber (8).
Herefter kan der foretages en tofaktor-ANOVA med koncentration og SentStadie og deres vekselvirkning med Rep(Konc) en vilkårlig faktor og Haletudse(Rep) en anden vilkårlig virkning.
Developmental stage data are used to determine if development is accelerated, asynchronous, delayed or unaffected.
Dette behandler stadig replikatet som analyseenheden og giver i al væsentlighed de samme resultater som en vægtet analyse af rep*sentstadie-gennemsnit, vægtet med antallet af dyr pr. gennemsnit.
Acceleration or delay of development is determined by making a comparison between the median stage achieved by the control and treated groups.
Hvis dataene ikke opfylder ANOVA's normalitets- eller varianshomogenitetskrav, kan der foretages en normaliseret rangordenstransformation for at fjerne denne indsigelse.
Asynchronous development is reported when the tissues examined are not malformed or abnormal, but the relative timing of the morphogenesis or development of different tissues is disrupted within a single tadpole.
Ud over standard-ANOVA F-test for virkningerne af Conc, SentStadie og vekselvirkningerne mellem dem, kan vekselvirknings-F-testen »deles« i to supplerende ANOVA F-test, en vedrørende de gennemsnitlige responser på tværs af koncentrationer for SentStadie=»Nej« og en anden vedrørende gennemsnitsresponserne på tværs af koncentrationer for SentStadie=»Ja«.
Differentiation and growth of the hind limbs are under control of thyroid hormones and are major developmental landmarks already used in the determination of developmental stage.
Yderligere sammenligninger af middelværdier for behandlingen i forhold til kontroller foretages inden for hvert enkelt SentStadie-niveau.
Hind limb development is used qualitatively in the determination of developmental stage, but is considered here as a quantitative endpoint.
En tendenstypeanalyse kan foretages ved hjælp af passende kontraster, eller der kan foretages simple parvise sammenligninger, hvis der er evidens for ikkemonoton dosisrespons inden for et niveau af SentStadie-variabel.
Therefore, hind limb length is measured as an endpoint to detect effects on the thyroid axis (Figure 2).
En Bonferroni-Holm-tilpasning til p-værdierne foretages kun, hvis den tilsvarende F-skive ikke er signifikant.
For consistency, hind limb length is measured on the left hind limb.
Dette kan gøres i SAS og — formentlig — andre statistiske softwarepakker.
Hind limb length is evaluated both at day 7 and at day 21 of the test.
Der kan opstå komplikationer, når der i nogle koncentrationer ikke er dyr i et sent stadium, men disse situationer kan håndteres på en enklere måde.
On day 7, measuring hind limb length is straightforward, as illustrated in Figure 2.
Testkemikalie et stof eller en blanding, som testes ved hjælp af denne testmetode.
However, measuring hind limb length on day 21 is more complicated due to bends in the limb.
COLLEMBOLFORMERINGSTEST I JORD
Therefore, measurements of hind limb length at day 21 should originate at the body wall and follow the midline of the limb through any angular deviations.
Denne testmetode svarer til OECD Test Guideline (TG) 232 (2009).
Changes in hind limb length at day 7, even if not evident at day 21, are still considered significant for potential thyroid activity.
Denne testmetode er designet til at vurdere virkningerne af kemikalier på collembolers (springhalers) formeringsevne i jord.
Length measurements are acquired from digital photographs using image analysis software as described in the OECD Guidance Document on Amphibian Thyroid Histology (9).
Den er baseret på eksisterende procedurer (1) (2).
Body Length and Wet Weight
Den partenogenetiske Folsomia candida og seksuelt reproducerende Folsomia fimetaria er to af de mest tilgængelige collembolarter, og de kan dyrkes og kan fås i handelen.
Determinations of snout to vent length (SVL) (Figure 2) and wet weight are included in the test protocol to assess possible effects of test chemicals on the growth rate of tadpoles in comparison to the control group and are useful in detecting generalised toxicity to the test chemical.
Når specifikke habitater, som ikke er dækket af de to arter, skal vurderes, kan fremgangsmåden også udvides til andre collembolarter, hvis de kan opfylde testens validitetskriterier.
Because the removal of adherent water for weight determinations can cause stressful conditions for tadpoles and may cause skin damage, these measurements are performed on the day 7 sub-sampled tadpoles and all remaining tadpoles at test termination (day 21).
Jordboende collemboler er økologisk relevante arter til økotoksikologiske test.
For consistency, use the cranial aspect of the vent as the caudal limit of the measurement.
Collemboler er heksapoder med et tyndt luft- og vandgennemtrængeligt exoskelet og repræsenterer leddyrarter med en anden eksponeringsvej og grad sammenlignet med regnorme og enchytraeider.
Snout to vent length (SVL) is used to assess tadpole growth as illustrated in Figure 2.
Collembolers populationstætheder når i almindelighed op på 105 m– 2 i jord- og førnlag i mange terrestriske økosystemer (3) (4).
Types of body length measurements and (B) Hind limb length measurements for X. laevis tadpoles (1)
Voksne individer måler typisk 0,5-5 mm, og deres bidrag til den samlede jordbaserede animalske biomasse og respiration er lavt, anslået 1-5 % (5).
While developmental stage and hind limb length are important endpoints to evaluate exposure-related changes in metamorphic development, developmental delay cannot, by itself, be considered a diagnostic indicator of anti-thyroidal activity.
Deres vigtigste rolle kan derfor være som potentielle regulatorer af processer gennem mikrobe- og mikrofaunaprædation.
Some changes may only be observable by routine histopathological analysis.
Springhaler er byttedyr for en bred vifte af hvirvelløse dyr under og over jorden såsom mider, tusindben, edderkopper, løbebiller (Carabidae) og rovbiller.
Diagnostic criteria include thyroid gland hypertrophy/atrophy, follicular cell hypertrophy, follicular cell hyperplasia, and as additional qualitative criteria:
Collemboler bidrager til nedbrydningsprocesser i sur jordbund, hvor de kan være de vigtigste jordlevende hvirvelløse dyr ud over enchytraeider, eftersom regnorme og diplopoder normalt er fraværende.
follicular lumen area, colloid quality and follicular cell height/shape.
F. fimetaria findes i hele verden og er almindelig i flere jordtyper fra sandede til lerede jorde og fra muld- til morjorde.
Severity grading (4 grades) should be reported.
Det er en øjeløs upigmenteret collembol.
Information on obtaining and processing samples for histological analysis and for performing histologic analyses on tissue samples is available in “Amphibian Metamorphosis Assay: Part 1 — Technical guidance for morphologic sampling and histological preparation” and “Amphibian Metamorphosis Assay: Part 2 — Approach to reading studies, diagnostic criteria, severity grading and atlas” (9).