natgillin/translations-raw huggingface dataset:
The FSDT validation phases (1 and 2) did cover oestrogenic, androgenic and steroidogenesis inhibiting chemicals.
a halak szokatlan reakcióinak előfordulási gyakorisága és a vizsgált vegyi anyag által kiváltott minden látható hatás.
The effects in the FSDT of oestrogen- and androgen antagonists can be seen in Table 1 but these MOA are less documented at present time.
Az eredményeket átlag ± szórás vagy standard hiba formában kell bemutatni.
In the test, fish are exposed, from newly fertilised egg until the completion of sexual differentiation, to at least three concentrations of the test chemical dissolved in water.
A minimálisan jegyzőkönyvezendő statisztikák a NOEC és a LOEC, illetve a konfidenciaintervallumok.
The test conditions should be flow-through unless not possible due to the availability or nature (e.g. limited solubility) of the test chemical.
A statisztikai folyamatábra (8. függelék) szerint kell eljárni.
The test starts with the placing of newly fertilised eggs (before cleavage of the blastodisc) in the test chambers.
Sumpter, 1996, »Inhibition of testicular growth in rainbow trout (Oncorhynchus mykiss) exposed to estrogenic alkylphenolic chemicals«, Environmental Toxicology and Chemistry 15, pp.
The loading of the chambers is described for each species in paragraph 27.
194-202.
For the validated fish species, Japanese medaka, the three-spined stickleback and zebrafish, the test is terminated at 60 dph.
Sumpter, J.P. and S. Jobling, 1995, »Vitellogenesis As A Biomarker for Estrogenic Contamination of the Aquatic Environment«, Environmental Health Perspectives 103, pp.
At test termination, all fish are euthanised humanely.
173-178.
A biological sample (blood plasma, liver or head/tail homogenate) is collected for VTG analysis from each fish and the remaining part is fixed for histological evaluation of the gonads to determine the phenotypic sex;
Hutchinson, S. Maddix, and H. Trip (1999), »An in vivo testing system for endocrine disruptors in fish early life stages using induction of vitellogenin«, Environmental Toxicology and Chemistry 18, pp.
optionally, histopathology (e.g. staging of gonads, severity of intersex) can be performed.
337-347.
A biological sample (the anal- or the dorsal fin) for the determination of the genetic sex is taken in species possessing appropriate markers (Appendices 9 and 10).
Pedersen, and P. Bjerregaard (2001a), »Development of an ELISA for vitellogenin in whole body homogenate of zebrafish (Danio rerio)«, Comparative Biochemistry and Physiology C-Toxicology & Pharmacology 130, pp.
An overview of relevant test conditions specific for validated species:
119-131.
Japanese medaka, the three-spined stickleback and zebrafish is provided in Appendix 2.
Andersen, L., P. Bjerregaard, and B. Korsgaard ( 2003), »Vitellogenin induction and brain aromatase activity in adult male and female zebrafish exposed to endocrine disrupters«, Fish Physiology and Biochemistry 28, pp.
Results from an acute toxicity test or other short-term toxicity assay [e.g. test method C.14 (34) and OECD TG 210 (1)], preferably performed with the species chosen for this test, should be available.
319-321.
This implies that the water solubility and the vapour pressure of the test chemical are known and a reliable analytical method for the quantification of the chemical in the test chambers, with known and reported accuracy and limit of detection, is available.
Petersen (2003), »Gonad development and vitellogenin production in zebrafish (Danio rerio) exposed to ethinylestradiol and methyltestosterone«, Aquatic Toxicology 65, pp.
Other useful information includes the structural formula, purity of the chemical, stability in water and light, pKa, Pow and results of a test for ready biodegradability (Test Method C.4) (35).
397-411.
Test acceptance criteria
Tyler (2002), »Utility of a juvenile fathead minnow screening assay for detecting (anti-)estrogenic substances«, Environmental Toxicology and Chemistry 21, pp.
The dissolved oxygen concentration should be at least 60 per cent of the air saturation value (ASV) throughout the test;
319-326.
The water temperature should not differ by more than ± 1,5 °C between test chambers at any one time during the exposure period and be maintained within the temperature ranges specified for the test species (Appendix 2);
Wang (2007), »Toxicity of the aromatase inhibitor letrozole to Japanese medaka (Oryzias latipes) eggs, larvae and breeding adults«, Comparative Biochemistry and Physiology C-Toxicology & Pharmacology 145, pp.
A validated method for analysis of the exposure chemical with a detection limit well below the lowest nominal concentration should be available and evidence should be gathered to demonstrate that the concentrations of the test chemical in solution have been satisfactorily maintained within ± 20 % of the mean measured values;
533-541.
Overall survival of fertilised eggs in the controls and, where relevant, in the solvent controls, should be greater than or equal to the limits defined in Appendix 2;
LeBlanc, and C.V.Sullivan (1999), »Fathead minnow (Pimephales promelas) vitellogenin: purification, characterization and quantitative immunoassay for the detection of estrogenic compounds«, Comparative Biochemistry and Physiology C-Toxicology & Pharmacology 123, pp.
Acceptance criteria related to growth and proportions of sex at termination of the test are based on data from the control groups (pooled solvent and water control unless they are significantly different, then solvent only):
113-125.
Japanese medaka
Porcher (2002), »Development and validation of an enzyme-linked immunosorbent assay to measure vitellogenin in the zebrafish (Danio rerio)«, Environmental Toxicology and Chemistry 21, pp.
Three-spined stickleback
1699-1708.
Growth
Hatano, H. Mizukami, M. Yamashita, R. Sakakibara, and E. Tamiya (2002), »Development and application of a monoclonal antibody-based sandwich ELISA for quantification of Japanese medaka (Oryzias latipes) vitellogenin«, Comparative Biochemistry and Physiology C-Toxicology & Pharmacology 132, pp.
Fish wet weight, blotted dry
161-169.
> 75 mg
Bengtsson (2004), »The juvenile three-spined stickleback (Gasterosteus aculeatus L.) as a model organism for endocrine disruption – II – kidney hypertrophy, vitellogenin and spiggin induction«, Aquatic Toxicology 70, pp.
Length (standard length)
311-326.
> 20 mm
Tatarazako, N., M. Koshio, H. Hori, M. Morita, and T. Iguchi (2004), »Validation of an enzyme-linked immunosorbent assay method for vitellogenin in the medaka«, Journal of Health Science 50, pp.
> 14 mm
301-308.
Sex ratio (% males or females)
Eidem, J.K., H. Kleivdal, K. Kroll, N. Denslow, R. van Aerle, C. Tyler, G. Panter, T. Hutchinson, and A. Goksoyr (2006), »Development and validation of a direct homologous quantitative sandwich ELISA for fathead minnow (Pimephales promelas) vitellogenin.
When a solvent is used it should have no statistical significant effect on survival and should not produce any endocrine disrupting effects or other adverse effects on the early-life stages as revealed by a solvent control.
Aquatic Toxicology«, 78, pp.
If a deviation from the test acceptance criteria is observed, the consequences should be considered in relation to the reliability of the test data and these considerations should be included in the reporting.
202-206.
Test chambers
Ankley (2006), »Evaluation of a commercial kit for measuring vitellogenin in the fathead minnow (Pimephales promelas)«, Ecotoxicology and Environmental Safety 64, pp.
Any glass, stainless steel or other chemically inert chambers can be used.
101-105.
The dimensions of the chambers should be large enough to allow compliance with loading rate criteria given below.
Holbech, H., Petersen, G. I., Norman, A., Örn, S, Norrgren, L., and Bjerregaard, P (2001b), »Suitability of zebrafish as test organism for detection of endocrine disrupting chemicals.
It is desirable that test chambers be randomly positioned in the test area.
Comparison of vitellogenin in plasma and whole body homogenate from zebrafish (Danio rerio) and rainbow trout (Oncorhynchus mykiss)«, Nordic Council of Ministers, TemaNord 2001:597, pp.
A randomised block design with each concentration being present in each block is preferable to a completely randomised design.
48-51.
The test chambers should be shielded from unwanted disturbance.
Porcher, and A. Goksoyr (2004), »Development of quantitative vitellogenin-ELISAs for fish test species used in endocrine disruptor screening«, Analytical and Bioanalytical Chemistry 378, pp.
Recommended fish species are given in Appendix 2.
621-633.
The procedures for inclusion of new species are given in paragraph 2.
Orn, S., S. Yamani, and L. Norrgren (2006), »Comparison of vitellogenin induction, sex ratio, and gonad morphology between zebrafish and Japanese medaka after exposure to 17 alpha-ethinylestradiol and 17 beta-trenbolone«, Archives of Environmental Contamination and Toxicology 51, pp.
Holding of parental fish
237-243.
Details on holding the parental fish under satisfactory conditions may be found in OECD TG 210(1).
Scholz, S. and N. Kluver (2009), »Effects of Endocrine Disrupters on Sexual, Gonadal Development in Fish, Sexual Development 3«, pp.
Parental fish should be fed once or twice a day with appropriate food.
136-151.
Handling of embryos and larvae
Fenske, M., G. Maack, C. Schafers, and H. Segner (2005), »An environmentally relevant concentration of estrogen induces arrest of male gonad development in zebrafish, Danio rerio«, Environmental Toxicology and Chemistry 24, pp.
Initially, embryos and larvae may be exposed within a main chamber in smaller glass or stainless steel chambers, fitted with mesh sides or ends to permit a flow of test chemical through the chamber.
1088-1098.
Non-turbulent flow through these small chambers may be induced by suspending them from an arm arranged to move the chamber up and down but always keeping the organisms submerged.
Nagahama (2004), »Two DM domain genes, DMY and DMRT1, involved in testicular differentiation and development in the medaka, Oryzias latipes«, Developmental Dynamics 231, pp.
Where egg containers, grids or meshes have been used to hold eggs within the main test chamber, these restraints should be removed after the larvae hatch, except that meshes should be retained to prevent the escape of the fish.
518-526.
If there is a need to transfer the larvae, they should not be exposed to the air and nets should not be used to release fish from egg containers.
Shinomiya, A., H. Otake, K. Togashi, S. Hamaguchi, and M. Sakaizumi (2004), »Field survey of sex-reversals in the medaka, Oryzias latipes: genotypic sexing of wild populations«, Zoological Science 21, pp.
The timing of this transfer varies with the species and transfer may not always be necessary.
613-619.
Any water in which the test species shows control survival at least as good as in water described in Appendix 3 is suitable as test water.
Flick (2007), »Collapse of a fish population after exposure to a synthetic estrogen«, Proceedings of the National Academy of Sciences of the United States of America 104, pp.
In order to ensure that the dilution water will not unduly influence the test result (for example by reacting with the test chemical) or adversely affect the performance of the brood stock, samples should be taken at intervals for analysis.
8897-8901.
Total organic carbon, conductivity, pH and suspended solids should be measured, for example every three months where dilution water is known to be relatively constant in quality.
Kidd (2009), »Interspecies differences in biochemical, histopathological, and population responses in four wild fish species exposed to ethynylestradiol added to a whole lake«, Canadian Journal of Fisheries and Aquatic Sciences 66, pp.
Measurements of heavy metals (e.g. Cu, Pb, Zn, Hg, Cd, Ni), major anions and cations (e.g. Ca2+, Mg2+, Na+, K+, Cl–, SO42–) and pesticides should be done, if water quality is questionable.
1920-1935.
Details about chemical analysis and water collection can be found in paragraph 34.
Tyler (2006), »Development of chronic tests for endocrine active chemicals – Part 1.
Flow-through system should be used if practically possible.
An extended fish early-life stage test for oestrogenic active chemicals in the fathead minnow (Pimephales promelas)«, Aquatic Toxicology 77, pp.
For flow-through tests, a system that continually dispenses and dilutes a stock solution of the test chemical (e.g. metering pump, proportional diluter, and saturator system) is necessary to deliver a series of concentrations to the test chambers.
279-290.
The flow rates of stock solutions and dilution water should be checked at intervals during the test and should not vary by more than 10 % throughout the test.
Petersen, P. Jackson, K. Hylland, L. Norrgren, and P. Bjerregaard (2006), »Detection of endocrine disrupters: Evaluation of a Fish Sexual Development Test (FSDT)«, Comparative Biochemistry and Physiology C-Toxicology & Pharmacology 144, pp.
A flow rate equivalent to at least five test chamber volumes per 24 hours has been found suitable (1).
57-66.
Care should be taken to avoid the use of plastic tubing or other materials, some of which may contain biologically active chemicals or may adsorb the test chemical.
Andersen, L., K. Kinnberg, H. Holbech, B. Korsgaard, and P. Bjerregaard (2004), »Evaluation of a 40 day assay for testing endocrine disrupters: Effects of an anti-estrogen and an aromatase inhibitor on sex ratio and vitellogenin concentrations in juvenile zebrafish (Danio rerio)«, Fish Physiology and Biochemistry 30, pp.
The stock solution should preferably be prepared without the use of solvents by simply mixing or agitating the test chemical in the dilution water by using mechanical means (e.g. stirring or ultrasonication).
257-266.
If the test chemical is difficult to dissolve in water, procedures described in the OECD Guidance Document on aquatic toxicity testing of difficult substances and mixtures should be followed (36).
Morthorst, J.E., H. Holbech, and P. Bjerregaard (2010), »Trenbolone causes irreversible masculinization of zebrafish at environmentally relevant concentrations«, Aquatic Toxicology 98, pp.
The use of solvents should be avoided but may be necessary in some cases in order to produce a suitably concentrated stock solution.
336-343.
Examples of suitable solvents are given in (36).
Metcalf (2003), »Effects of the antiandrogens, vinclozolin and cyproterone acetate on gonadal development in the Japanese medaka (Oryzias latipes)«, Aquatic Toxicology 63, pp.
Semi-static test conditions should be avoided unless justification is provided on compelling reasons associated with the test chemical (e.g. stability, limited availability, high cost or hazard).
391-403.
For the semi-static technique, two different renewal procedures may be followed.
Tyler (2004), »Successful detection of (anti-) androgenic and aromatase inhibitors in pre-spawning adult fathead minnows (Pimephales promelas) using easily measured endpoints of sexual development«, Aquatic Toxicology 70, pp.
Either new test solutions are prepared in clean chambers and surviving eggs and larvae gently transferred into the new chambers, or the test organisms are retained in the test chambers whilst a proportion (at least two thirds) of the test water is changed daily.
11-21.
Conditions of Exposure
Petersen, and P. Bjerregaard (2007), »Effects of the fungicide prochloraz on the sexual development of zebrafish (Danio rerio)«, Comparative Biochemistry and Physiology C-Toxicology & Pharmacology 145, pp.
Collection of eggs and duration
165-170.
To avoid genetic bias, eggs are collected from a minimum of three breeding pairs or groups, mixed and randomly selected to initiate the test.
E melléklet C.14. fejezete: Halivadékok növekedési vizsgálata.
For the three-spined stickleback, see the description of artificial fertilisation in Appendix 11.
E melléklet C.4. fejezete: Gyors biológiai lebonthatóság.
The test should start as soon as possible after the eggs have been fertilised, the embryos preferably being immersed in the test solutions before cleavage of the blastodisc commences, or as close as possible after this stage and no later than 12 h post fertilisation.
E melléklet C.37. fejezete: 21 napos halvizsgálat.
The test should continue until sexual differentiation in the control group is completed (60 dph for Japanese medaka, the three-spined stickleback and zebrafish).
Az ösztrogén- és androgénhatás, valamint az aromatázgátlás rövid távú szűrővizsgálata.
Loading
Schäfers, C., Teigeler, M., Wenzel, A., Maack, G., Fenske, M., Segner, H (2007), »Concentration- and time-dependent effects of the synthetic estrogen, 17 alpha-ethinylestradiol, on reproductive capabilities of the zebrafish, Danio rerio« Journal of Toxicology and Environmental Health-Part A, 70, 9-10 pp 768-779.
The number of fertilised eggs at the start of the test should be at least 120 per concentration divided between a minimum of 4 replicates (square root allocation to control is accepted).
Rövidítések és fogalommeghatározások
The eggs should be randomly distributed (by using statistical tables for randomisation) among treatments.
Apikális végpontok
The loading rate (for definition, see Appendix 1) should be low enough in order that a dissolved oxygen concentration of at least 60 % of the ASV can be maintained without direct aeration of the chambers.
populációszinten okoznak hatást
For flow-through tests, a loading rate not exceeding 0,5 g/l per 24 hours, and not exceeding 5 g/l of solution at any time is recommended.
levegőtelítettségi érték
No later than 28 days post fertilisation the number of fish per replicate should be redistributed, so that each replicate contains as equal a number of fish as possible.
az egyedek szintjén okoz hatást
If exposure related mortality occurs, the number of replicates should be reduced appropriately so that fish density between treatment levels is kept as equal as possible.
anyag vagy keverék.
The photoperiod and water temperature should be appropriate for the test species (see Appendix 2 for experimental conditions for the FSDT).
a kikelés utáni napok száma
To obtain a sufficient growth rate, fish should be fed at least twice daily (accepting once daily on weekends), separated by at least three hours between each feed.
Hal tömege
Surplus food and faeces should be removed, as necessary, to avoid accumulation of waste.
a hal nedves tömege (szárazra itatva)
As experience is gained, food and feeding regimes are continuously being refined to improve survival and optimise growth.
halak ivari fejlődésének vizsgálata
Effort should therefore be made to confirm the proposed regime with acknowledged experts.
hypothalamus-hipofízis-gonád tengely
Feeding should be withheld 24 hours before ending the test.
Interszexuális hal
Examples of appropriate food items are listed in Appendix 2 (see also the OECD Fish Testing Framework (39).
olyan hal, amelyben a here hat elemzett metszetében több mint egy petesejt van jelen, vagy spermatogén sejtek vannak jelen a petefészekben (igen/nem)
Test chemicals should be spaced as described in Appendix 4.
a halak nedvesen mért tömege egységnyi mennyiségű vízben
A minimum of three test concentrations in at least four replicates should be used.
hatásmechanizmus
The curve relating LC50 to period of exposure in the acute studies available should be considered when selecting the range of test concentrations.
reverz transzkriptáz polimeráz láncreakció
Five test concentrations are recommended if the data are to be used for risk assessment.
Differenciálatlan hal
Concentrations of the chemical higher than 10 % of the acute adult LC50 or 10 mg/l, whichever is the lower, need not be tested.
olyan hal, amelynek ivarmirigyeiben nincsenek észlelhető csírasejtek.
The maximum test concentration should be 10 % of the LC50 on the larval/juvenile life-stage.
Az FSDT kísérleti körülményei (édesvízi fajok)
Controls
Japán fogasponty (Oryzias latipes)
A dilution water control (≥ 4 replicates) and, if relevant, a solvent control (≥ 4 replicates) should be run in addition to the test concentrations.
Zebradánió (Danio rerio)
Only solvents that have been investigated not to have any statistical significant influence on the test endpoints should be used in the test.
Tüskés pikó (Gasterostreus aculeatus)
Where a solvent is used, its final concentration should not be greater than 0,1 ml/l (36) and it should be the same concentration in all test chambers, except the dilution water control.
Átfolyásos vagy félstatikus
However, every effort should be made to avoid the use of such solvent or keep solvent's concentrations to a minimum.
Fluoreszcens izzók (széles spektrum)
Frequency of Analytical Determinations and Measurements
Fényerő
Chemical analysis of the test chemical concentration should be performed before initiation of the test to check compliance with the acceptance criteria.
10–20 μE/m2/s, 540–1080 lux vagy 50–100 ft-c (laboratóriumi környezeti értékek)
All replicates should be analysed individually at the beginning and termination of the test.
12–16 óra világos, 8–12 óra sötét
One replicate per test concentration should be analysed at least once per week during the test, changing systematically between replicates (1,2,3,4,1,2…).
Kamra minimális mérete
If samples are stored to be analysed at a later time, the storage method of the samples should be previously validated.
Az egyes kamráknak legalább 7 l vizet kell tartalmaznia
Samples should be filtered (e.g. using a 0,45 μm pore size) or centrifuged to ensure that the determinations are made on the chemical in true solution.
Vizsgálati organizmusok életkora az expozíció elején
During the test, dissolved oxygen, pH, total hardness, conductivity, salinity (if relevant), and temperature should be measured in all test chambers.
Újonnan megtermékenyített ikrák (korai hólyagcsíra stádium)
As a minimum dissolved oxygen, salinity (if relevant), and temperature should be measured weekly, and pH, conductivity and hardness at the beginning and at the end of the test.
Újonnan megtermékenyített ikrák
Temperature should preferably be monitored continuously in at least one test chamber.
Ikrák száma kezelésenként
Results should be based on measured concentrations.
Legalább 120
However, if the concentration of the test chemical in solution has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test, then the results can either be based on nominal or measured values.
Legalább 3 (plusz megfelelő kontrollok)
Observations and measurements
Ismétlések száma kezelésenként
Stage of embryonic development
Legalább 4 (kivéve, ha a kontrollokat négyzetgyökösen allokálják)
The exposure should begin as soon as possible after fertilisation and before cleavage of the blastodisc commences and no later than 12 h post fertilisation to ensure exposure during early embryonic development.
Élő Artemia, fagyasztott felnőtt sósvízi rák, pelyhesített táplálék stb.
Hatching and survival
Naponta kétszeri etetés javasolt.
Observations on hatching and survival should be made at least once daily and numbers recorded.
Különleges sült táplálék, élő Artemia, fagyasztott felnőtt sósvízi rák, pelyhesített táplálék stb.
Dead embryos, larvae and juvenile fish should be removed as soon as observed since they can decompose rapidly and may be broken up by the actions of the other fish.
Naponta kétszeri etetés javasolt.
Extreme care should be taken when removing dead individuals not to knock or physically damage adjacent eggs/larvae, these being extremely delicate and sensitive.
Nincs, kivéve ha az oldott oxigénkoncentráció a 60 %-os telítettség alá esik
Criteria for death vary according to life stage:
Nincs, kivéve ha az oldott oxigénkoncentráció a 70 %-os telítettség alá esik
for eggs:
Tiszta felszíni víz, kútvíz vagy mesterséges víz
particularly in the early stages, a marked loss of translucency and change in coloration, caused by coagulation and/or precipitation of protein, leading to a white opaque appearance;
Vizsgált vegyi anyaggal történő expozíció időtartama
for larvae and juvenile fish:
kikelés eredményessége, túlélési arány, általános morfológia, VTG ivarmirigy-szövettan, genetikai ivar ivararány