natgillin/translations-raw huggingface dataset:
Použije-li se Dunnettův test, pro další pokračování není třeba provádět ANOVA F-test významnosti.
E tube (plastic or glass, inner diameter ca.
Pro podrobnosti viz schéma v dodatku 8.
8 mm) to collect adult midges from vessel
Výsledky by měly být uvedeny v tabulkách jako střední hodnoty u samečků, samiček, intersexuálů a nediferencovaných jedinců odděleně pro každou koncentraci.
Schematic presentation of a life-cycle test:
Měla by být zdůrazněna statistická významnost u fenotypových samiček a fenotypových samečků.
A 1st generation — test vessels containing a sediment-water system, eight replicates, 20 first instar larvae per vessel
Zkoušená chemická látka: skutečná koncentrace
B four test vessels for each breeding cage, A and B
Skutečné koncentrace zkoušené chemické látky v nádržích by měly být analyzovány s četností popsanou v odstavci 34.
C breeding cages (A and B) for swarming, mating and oviposition
Výsledky by měly být uvedeny v tabulkách jako střední koncentrace ± směrodatná odchylka na bázi opakování a rovněž i na bázi koncentrací spolu s informací o počtu vzorků a zdůrazní se odlehlé hodnoty od střední zkušební koncentrace ± 20 %.
D crystallising dishes for deposition of egg ropes
Příklady obsahuje validační zpráva pro fázi 2 zkoušky FSDT (42).
E micro plates, one well for each egg rope
V případě, že se naměřené koncentrace zkoušené chemické látky ve zkušebních roztocích pohybují na úrovních blízkých mezi stanovitelnosti dané analytické metody, by výsledky měly být interpretovány opatrně.
F 2nd generation — test vessels containing a sediment-water system, eight replicates, 20 first instar larvae per vessel
Použitý zkušební postup (např. průtokový, semistatický / s obnovováním);
C.41.
uspořádání zkoušky včetně zkušebních koncentrací, metoda přípravy zásobních roztoků (v příloze), četnost obnovování (je-li použito, uvede se solubilizační činidlo a jeho koncentrace),
FISH SEXUAL DEVELOPMENT TEST
nominální zkušební koncentrace, průměry naměřených hodnot v jednotlivých zkušebních nádržích, jejich směrodatné odchylky a metody jejich stanovení (použitá analytická metoda by měla být uvedena v příloze); důkazy o tom, že naměřené hodnoty odpovídají koncentracím zkoušené chemické látky ve skutečném roztoku,
This test method is equivalent to OECD test guideline (TG) 234 (2011).
kvalita vody ve zkušebních nádržích:
It is based on a decision from 1998 to develop new or update existing test methods for the screening and testing of potential endocrine disrupters.
podrobné informace o krmení (např. druh krmiva/krmiv, původ, podávané množství a frekvence krmení) a podle potřeby analýzy na přítomnost kontaminujících látek (např. polychlorovaných bifenylů, polyaromatických uhlovodíků a organochlorových pesticidů).
The Fish Sexual Development Test (FSDT) was identified as a promising test method covering a sensitive fish life stage responsive to both oestrogen and androgen-like chemicals.
Doklady o tom, že kontrolní skupiny splnily kritéria validity:
The test method went through an inter-laboratory validation exercise from 2006 to 2010, where Japanese medaka (Oryzias latipes), zebrafish (Danio rerio) and three spined stickleback (Gasterosteus aculeatus) were validated and fathead minnow (Pimephales promelas) was partially validated (41) (42) (43).
údaje o míře vylíhnutí by měly být uvedeny v tabulkách jako procentní podíl v každém opakování a pro každou koncentraci.
This protocol includes Japanese medaka, the three-spined stickleback and zebrafish.
Měly by být zdůrazněny odlehlé hodnoty od kritérií přijatelnosti (v kontrolních skupinách).
The protocol is in principle an enhancement of OECD TG 210 Fish, Early Life Stage Toxicity Test (1), where the exposure is continued until the fish are sexually differentiated, i.e. about 60 days post-hatch (dph) for Japanese medaka, the three-spined stickleback and zebrafish (the exposure period can be shorter or longer for other species that are validated in the future), and endocrine-sensitive endpoints are added.
Měly by být zdůrazněny odlehlé hodnoty od kritérií validity (v kontrolních skupinách),
The combination of the two core endocrine endpoints, vitellogenin (VTG) concentration and phenotypic sex ratio enable the test to indicate the mode of action of the test chemical.
jasné uvedení dosažených výsledků v jednotlivých sledovaných ukazatelích:
Due to the population-relevant change in phenotypic sex ratio, the FSDT can be used for hazard and risk assessment.
přežití embryí a úspěšnost líhnutí;
However, if the test is used for hazard or risk assessment, the stickleback should not be used because the validation data available so far showed that in this species the alterations of phenotypic sex ratio by the test chemicals were uncommon.
zevní odchylky;
The protocol is based on fish exposed via water to chemicals during the sex labile period in which the fish is expected to be most sensitive to the effects of endocrine disrupting chemicals that interfere with sexual development.
délka a hmotnost;
Two core endpoints are measured as indicators of endocrine-associated developmental aberrations, the VTG concentrations and sex ratios (proportions of sex) determined via gonad histology.
naměřené hodnoty VTG (v ng/g homogenátu, ng/ml plazmy nebo ng/mg jater);
Gonadal histopathology (evaluation and staging of oocytes and spermatogenetic cells) is optional.
histologie gonád, poměr pohlaví, údaje o genetickém pohlaví;
Additionally, the genetic sex is determined whenever possible (e.g. in Japanese medaka and the three spined stickleback).
výskyt jakýchkoli neobvyklých reakcí ryb a jakékoli viditelné účinky způsobené zkoušenou chemickou látkou.
The presence of a genetic sex marker is a considerable advantage as it increases the power of the sex ratio statistics and enables the detection of individual phenotypic sex reversal.
Výsledky by měly být uvedeny jako střední hodnoty ± směrodatná odchylka (SD) nebo střední směrodatná chyba (SE).
Other apical endpoints that should be measured include hatching rate, survival, length and body weight.
Statistické údaje by měly být uvedeny minimálně jako NOEC a LOEC a intervaly spolehlivosti.
The test method might be adaptable to other species than those mentioned above provided that the other species undergo a validation equal to the one accomplished for Japanese medaka, the three-spined stickleback and zebrafish, that the control fish are sexually differentiated at the end of the test, that VTG levels are sufficiently high to detect significant chemical-related variations, and that sensitivity of the test system is established using endocrine active reference chemicals ((anti)-oestrogens, (anti)-androgens, aromatase inhibitors etc).
Mělo by se postupovat podle vývojového diagramu pro provedení statistické analýzy (dodatek 8).
In addition, any validation report(s) referring to FSDT data using other species should be reviewed by the OECD, and the validation outcome should be considered as satisfactory.
194–202.
Initial considerations and limitations
Sumpter, J.
VTG is normally produced by the liver of female oviparous vertebrates in response to circulating endogenous oestrogen (2).
P. and Jobling, S.
The VTG synthesis is very limited, though detectable, in immature fish and adult male fish because they lack sufficient circulating oestrogen.
(1995) Vitellogenesis As A Biomarker for Estrogenic Contamination of the Aquatic Environment, Environmental Health Perspectives 103, pp.
However, the liver is capable of synthesising and secreting VTG in response to exogenous oestrogen stimulation (3) (4) (5).
173–178.
The measurement of VTG serves for the detection of chemicals with oestrogenic, anti-oestrogenic, androgenic modes of action and chemicals that interfere with steroidogenesis as for example aromatase inhibitors.
337–347.
The detection of oestrogenic chemicals is possible via the measurement of VTG induction in male fish, and it has been abundantly documented in the scientific peer-reviewed literature.
119–131.
VTG induction has also been demonstrated following exposure to aromatisable androgens (6) (7).
Andersen, L., Bjerregaard, P. and Korsgaard, B.
A reduction in the circulating level of oestrogen in females, for instance through the inhibition of the aromatase converting the endogenous androgen to the natural oestrogen 17β-oestradiol, causes a decrease in the VTG concentration, which is used to detect chemicals having aromatase inhibiting properties or steroidogenesis inhibitors more broadly (33).
(2003) Vitellogenin induction and brain aromatase activity in adult male and female zebrafish exposed to endocrine disrupters, Fish Physiology and Biochemistry 28, pp.
The biological relevance of the VTG response following oestrogenic/aromatase inhibition is established and has been broadly documented (8) (9).
319–321.
However, it is possible that production of VTG in females can also be affected by general toxicity and non-endocrine toxic modes of action.
397–411.
Several measurement methods have been successfully developed and standardised for routine use to quantify VTG in blood, liver, whole body or head/tail homogenate samples collected from individual fish.
319–326.
This is the case for zebrafish, three-spined stickleback and Japanese medaka and also the partially validated species fathead minnow;
533–541.
species-specific Enzyme-Linked Immunosorbent Assay (ELISA) methods using immunochemistry for the quantification of VTG are available (5) (10) (11) (12) (13) (14) (15) (16).
purification, characterization and quantitative immunoassay for the detection of estrogenic compounds, Comparative Biochemistry and Physiology C-Toxicology & Pharmacology 123, pp.
In Japanese medaka and zebrafish, there is a good correlation between VTG measured from blood plasma, liver and homogenate samples although homogenates tend to show slightly lower values than plasma (17) (18) (19).
113–125.
Appendix 5 provides the recommended procedures for sample collection for VTG analysis.
1699–1708.
Change in the phenotypic sex ratio (proportions of sex) is an endpoint reflecting sex reversal.
Hatano, H.
In principle, oestrogens, anti-oestrogens, androgens, anti-androgens and steroidogenesis inhibiting chemicals can affect the sex ratio of developing fish (20).
Mizukami, M.
It has been shown that this sex reversal is partly reversible in zebrafish (21) following oestrogen-like chemical exposure, whereas sex reversal following androgen-like chemical exposure is permanent (30).
Yamashita, R.
The sex is defined as female, male, intersex (both oocytes and spermatogenetic cells in one gonad) or undifferentiated, determined in individual fish via histological examination of the gonads.
Sakakibara, and E.
Guidance is given in Appendix 7 and in the OECD Guidance Document on the Diagnosis of Endocrine-Related Histopathology of Fish Gonads (22).
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.
Genetic sex is examined via genetic markers when they exist in a given fish species.
161–169.
In Japanese medaka the female XX or male XY genes can be detected by Polymerase Chain-Reaction (PCR), or the Y-linked DM domain gene (DMY) can be analysed (DMY negative or positive) as described in (23) (24).
311–326.
In three-spined stickleback, there is an equivalent PCR method for genetic sex determination described in Appendix 10.
Tatarazako, N., M.
Where the genetic sex can be individually linked to the phenotypic sex, the power of the test is improved and therefore genetic sex should be determined in species with documented genetic sex markers.
Koshio, H.
The two core endocrine endpoints, VTG and sex ratio, can in combination demonstrate the endocrine mode of action (MOA) of the chemical (Table 1).
Hori, M.
The sex ratio is a population relevant biomarker (25) (26) and for some well defined modes of action, the FSDT results may be used for hazard and risk assessment purposes when deemed appropriate by the regulatory agency.
Morita, and T.
These modes of action are at present oestrogens, androgens and steroidogenesis inhibitors.
Iguchi (2004) Validation of an enzyme-linked immunosorbent assay method for vitellogenin in the medaka, Journal of Health Science 50, pp.
Reaction of the endocrine endpoints to different modes of action of chemicals
301–308.
↑ = increasing, ↓ = decreasing, — = not investigated
Eidem, J.
References
K., H.
Weak oestrogen agonist
Kleivdal, K.
↑♀ or ↑Undiff
Kroll, N.
Strong oestrogen agonist
Denslow, R. van Aerle, C.
↑♀ or ↑Undiff, No ♂
Tyler, G.
Oestrogen antagonist
Panter, T.
↓♀, ↑Undiff.
Hutchinson, and A.
Androgen agonist
Goksoyr (2006) Development and validation of a direct homologous quantitative sandwich ELISA for fathead minnow (Pimephales promelas) vitellogenin.
↑♂, No ♀
Aquatic Toxicology, 78, pp.
Androgen antagonist
202–206.
↑Intersex
101–105.
Aromatase inhibitor
Holbech, H., Petersen, G.
The FSDT does not cover the reproductive life stage of the fish and therefore chemicals that are suspected to affect reproduction at lower concentrations than sexual development should be examined in a test that covers reproduction.
I., Norman, A., Örn, S, Norrgren, L., and Bjerregaard, P (2001b) Suitability of zebrafish as test organism for detection of endocrine disrupting chemicals.
Definitions for the purpose of this Test Method are given in Appendix 1.
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.
The in vivo FSDT is intended to detect chemicals with androgenic and oestrogenic properties as well as anti-androgenic, anti-oestrogenic and steroidogenesis inhibiting properties.
48–51.
The FSDT validation phases (1 and 2) did cover oestrogenic, androgenic and steroidogenesis inhibiting chemicals.
621–633.
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.
Orn, S., S.
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.
Yamani, and L.
The test conditions should be flow-through unless not possible due to the availability or nature (e.g. limited solubility) of the test chemical.
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.
The test starts with the placing of newly fertilised eggs (before cleavage of the blastodisc) in the test chambers.
237–243.
The loading of the chambers is described for each species in paragraph 27.
Scholz, S. and N.
For the validated fish species, Japanese medaka, the three-spined stickleback and zebrafish, the test is terminated at 60 dph.
Kluver (2009) Effects of Endocrine Disrupters on Sexual, Gonadal Development in Fish, Sexual Development 3, pp.
At test termination, all fish are euthanised humanely.
136–151.
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;
Fenske, M., G.
optionally, histopathology (e.g. staging of gonads, severity of intersex) can be performed.
Maack, C.
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).
Schafers, and H.
An overview of relevant test conditions specific for validated species:
Segner (2005) An environmentally relevant concentration of estrogen induces arrest of male gonad development in zebrafish, Danio rerio, Environmental Toxicology and Chemistry 24, pp.
Japanese medaka, the three-spined stickleback and zebrafish is provided in Appendix 2.
1088–1098.
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.
Nagahama (2004) Two DM domain genes, DMY and DMRT1, involved in testicular differentiation and development in the medaka, Oryzias latipes, Developmental Dynamics 231, pp.
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.
518–526.
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).
Shinomiya, A., H.
Test acceptance criteria
Otake, K.
The dissolved oxygen concentration should be at least 60 per cent of the air saturation value (ASV) throughout the test;
Togashi, S.
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);
Hamaguchi, and M.
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;
Sakaizumi (2004) Field survey of sex-reversals in the medaka, Oryzias latipes: genotypic sexing of wild populations, Zoological Science 21, pp.
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;
613–619.
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):
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.
Japanese medaka
8897–8901.
Three-spined stickleback
P., R.
Growth
E.
Fish wet weight, blotted dry
Evans, K.
> 75 mg
G.
Length (standard length)
Wautier, K.
> 20 mm
H.
> 14 mm
Mills, P.
Sex ratio (% males or females)
J.
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.
Blanchfield, B.
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.
J.
Test chambers
Park, C.
Any glass, stainless steel or other chemically inert chambers can be used.
L.
The dimensions of the chambers should be large enough to allow compliance with loading rate criteria given below.
Baron, and K.
It is desirable that test chambers be randomly positioned in the test area.
A.
A randomised block design with each concentration being present in each block is preferable to a completely randomised design.
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.
The test chambers should be shielded from unwanted disturbance.
1920–1935.
Recommended fish species are given in Appendix 2.
An extended fish early-life stage test for oestrogenic active chemicals in the fathead minnow (Pimephales promelas), Aquatic Toxicology 77, pp.
The procedures for inclusion of new species are given in paragraph 2.
279–290.
Holding of parental fish
Evaluation of a Fish Sexual Development Test (FSDT), Comparative Biochemistry and Physiology C-Toxicology & Pharmacology 144, pp.
Details on holding the parental fish under satisfactory conditions may be found in OECD TG 210(1).
57–66.
Parental fish should be fed once or twice a day with appropriate food.
Andersen, L., K.
Handling of embryos and larvae
Kinnberg, H.
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.
Holbech, B.
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.
Korsgaard, and P.
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.
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.
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.
257–266.
The timing of this transfer varies with the species and transfer may not always be necessary.
Morthorst, J.
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.
E., H.
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.
Holbech, and P.
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.
Bjerregaard (2010) Trenbolone causes irreversible masculinization of zebrafish at environmentally relevant concentrations, Aquatic Toxicology 98, 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.
336–343.
Details about chemical analysis and water collection can be found in paragraph 34.
Metcalf (2003) ‚Effects of the antiandrogens, vinclozolin and cyproterone acetate on gonadal development in the Japanese medaka (Oryzias latipes)‘, Aquatic Toxicology 63, pp.
Flow-through system should be used if practically possible.
391–403.
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.
11–21.
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.
165–170.
A flow rate equivalent to at least five test chamber volumes per 24 hours has been found suitable (1).
Kapitola C.14 této přílohy, Růstová zkouška na nedospělých rybách.
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.
Kapitola C.4 této přílohy, Stanovení snadné biologické rozložitelnosti.
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).
Kapitola C.37 této přílohy, 21denní zkouška na rybách:
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).
krátkodobý screening estrogenní a androgenní aktivity a inhibice aromatázy.
The use of solvents should be avoided but may be necessary in some cases in order to produce a suitably concentrated stock solution.