natgillin/translations-raw huggingface dataset:
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).
datele privind rata de eclozare trebuie prezentate în tabele ca procente per replică și per concentrație.
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).
Valorile aberante din criteriile de acceptanță (în controale) trebuie evidențiate.
Appendix 5 provides the recommended procedures for sample collection for VTG analysis.
Supraviețuirea trebuie prezentată ca procent per replică și per concentrație.
Change in the phenotypic sex ratio (proportions of sex) is an endpoint reflecting sex reversal.
Valorile aberante din criteriile de valabilitate (în controale) trebuie evidențiate;
In principle, oestrogens, anti-oestrogens, androgens, anti-androgens and steroidogenesis inhibiting chemicals can affect the sex ratio of developing fish (20).
Indicarea clară a rezultatelor obținute privind diferiții parametri studiați observați:
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).
supraviețuire embrionilor și succesul eclozării;
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.
anomalii externe;
Guidance is given in Appendix 7 and in the OECD Guidance Document on the Diagnosis of Endocrine-Related Histopathology of Fish Gonads (22).
lungime și greutate;
Genetic sex is examined via genetic markers when they exist in a given fish species.
măsurători ale VTG (ng/g omogenat, ng/ml plasmă sau ng/mg ficat);
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).
histologia gonadelor, raportul sexelor, datele privind sexul genetic;
In three-spined stickleback, there is an equivalent PCR method for genetic sex determination described in Appendix 10.
incidența reacțiilor neobișnuite per pește și orice efect vizibil produs de substanța chimică testată.
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.
Rezultatele trebuie prezentate ca valori medii ± deviație standard (standard deviation – SD) sau eroare standard (standard error – SE).
The two core endocrine endpoints, VTG and sex ratio, can in combination demonstrate the endocrine mode of action (MOA) of the chemical (Table 1).
Statisticile trebuie raportate minimum ca NOEC și LOEC și intervale de încredere.
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.
Organigrama statistică (apendicele 8) trebuie urmată.
These modes of action are at present oestrogens, androgens and steroidogenesis inhibitors.
Sumpter, 1996, «Inhibition of testicular growth in rainbow trout (Oncorhynchus mykiss) exposed to estrogenic alkylphenolic chemicals», Environmental Toxicology and Chemistry 15, pp.
Reaction of the endocrine endpoints to different modes of action of chemicals
194-202.
↑ = increasing, ↓ = decreasing, — = not investigated
Sumpter, J.P. and S. Jobling, 1995, «Vitellogenesis As A Biomarker for Estrogenic Contamination of the Aquatic Environment», Environmental Health Perspectives 103, pp.
References
173-178.
Weak oestrogen agonist
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.
↑♀ or ↑Undiff
337-347.
Strong oestrogen agonist
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.
↑♀ or ↑Undiff, No ♂
119-131.
Oestrogen antagonist
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.
↓♀, ↑Undiff.
319-321.
Androgen agonist
Petersen (2003), «Gonad development and vitellogenin production in zebrafish (Danio rerio) exposed to ethinylestradiol and methyltestosterone», Aquatic Toxicology 65, pp.
↑♂, No ♀
397-411.
Androgen antagonist
Tyler (2002), «Utility of a juvenile fathead minnow screening assay for detecting (anti-)estrogenic substances», Environmental Toxicology and Chemistry 21, pp.
↑Intersex
319-326.
Aromatase inhibitor
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.
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.
533-541.
Definitions for the purpose of this Test Method are given in Appendix 1.
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.
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.
113-125.
The FSDT validation phases (1 and 2) did cover oestrogenic, androgenic and steroidogenesis inhibiting chemicals.
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.
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.
1699-1708.
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.
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.
The test conditions should be flow-through unless not possible due to the availability or nature (e.g. limited solubility) of the test chemical.
161-169.
The test starts with the placing of newly fertilised eggs (before cleavage of the blastodisc) in the test chambers.
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.
The loading of the chambers is described for each species in paragraph 27.
311-326.
For the validated fish species, Japanese medaka, the three-spined stickleback and zebrafish, the test is terminated at 60 dph.
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.
At test termination, all fish are euthanised humanely.
301-308.
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;
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.
optionally, histopathology (e.g. staging of gonads, severity of intersex) can be performed.
Aquatic Toxicology», 78, pp.
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).
202-206.
An overview of relevant test conditions specific for validated species:
Ankley (2006), «Evaluation of a commercial kit for measuring vitellogenin in the fathead minnow (Pimephales promelas)», Ecotoxicology and Environmental Safety 64, pp.
Japanese medaka, the three-spined stickleback and zebrafish is provided in Appendix 2.
101-105.
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.
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.
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.
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.
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).
48-51.
Test acceptance criteria
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.
The dissolved oxygen concentration should be at least 60 per cent of the air saturation value (ASV) throughout the test;
621-633.
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);
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.
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;
237-243.
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;
Scholz, S. and N. Kluver (2009), «Effects of Endocrine Disrupters on Sexual, Gonadal Development in Fish, Sexual Development 3», 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):
136-151.
Japanese medaka
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.
Three-spined stickleback
1088-1098.
Growth
Nagahama (2004), «Two DM domain genes, DMY and DMRT1, involved in testicular differentiation and development in the medaka, Oryzias latipes», Developmental Dynamics 231, pp.
Fish wet weight, blotted dry
518-526.
> 75 mg
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.
Length (standard length)
613-619.
> 20 mm
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.
> 14 mm
8897-8901.
Sex ratio (% males or females)
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.
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.
1920-1935.
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.
Tyler (2006), «Development of chronic tests for endocrine active chemicals – Part 1.
Test chambers
An extended fish early-life stage test for oestrogenic active chemicals in the fathead minnow (Pimephales promelas)», Aquatic Toxicology 77, pp.
Any glass, stainless steel or other chemically inert chambers can be used.
279-290.
The dimensions of the chambers should be large enough to allow compliance with loading rate criteria given below.
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.
It is desirable that test chambers be randomly positioned in the test area.
57-66.
A randomised block design with each concentration being present in each block is preferable to a completely randomised design.
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 test chambers should be shielded from unwanted disturbance.
257-266.
Recommended fish species are given in Appendix 2.
Morthorst, J.E., H. Holbech, and P. Bjerregaard (2010), «Trenbolone causes irreversible masculinization of zebrafish at environmentally relevant concentrations», Aquatic Toxicology 98, pp.
The procedures for inclusion of new species are given in paragraph 2.
336-343.
Holding of parental fish
Metcalf (2003), «Effects of the antiandrogens, vinclozolin and cyproterone acetate on gonadal development in the Japanese medaka (Oryzias latipes)», Aquatic Toxicology 63, pp.
Details on holding the parental fish under satisfactory conditions may be found in OECD TG 210(1).
391-403.
Parental fish should be fed once or twice a day with appropriate food.
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.
Handling of embryos and larvae
11-21.
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.
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.
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.
165-170.
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.
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.
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.
Abrevieri și definiții
The timing of this transfer varies with the species and transfer may not always be necessary.
Parametrii studiați apicali
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.
care cauzează efecte la nivelul populației
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.
Valoarea de saturație din aer (Air saturation value)
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.
Care cauzează efecte la nivel individual
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.
o substanță sau un amestec.
Details about chemical analysis and water collection can be found in paragraph 34.
Zile de la eclozare (Days post hatch)
Flow-through system should be used if practically possible.
Gena domeniului DM Y-specifică necesară pentru dezvoltarea masculilor din specia Oryzias latipes
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.
Test de imunoadsorbție cu anticorpi marcați enzimatic
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.
Greutate pește
A flow rate equivalent to at least five test chamber volumes per 24 hours has been found suitable (1).
Greutatea umedă peștilor (uscați prin tamponare)
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.
Test de dezvoltare sexuală a peștilor (Fish Sexual Development Test)
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).
Axul hipotalamo-hipofizo-gonadal.
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).
Pești hermafrodiți
The use of solvents should be avoided but may be necessary in some cases in order to produce a suitably concentrated stock solution.
Pești care prezintă mai multe ovocite în testicul pe 6 secțiuni analizate sau celule spermatogenetice în ovare (da/nu)
Examples of suitable solvents are given in (36).
Greutatea umedă a unui pește per volum de apă
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).
Mod de acțiune
For the semi-static technique, two different renewal procedures may be followed.
Reacție de polimerizare în lanț a revers transcriptazei
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.
Pești nediferențiați
Conditions of Exposure
Pești cu gonade care gonade care nu prezintă celulele germinale vizibile.
Collection of eggs and duration
Vitelogenină
To avoid genetic bias, eggs are collected from a minimum of three breeding pairs or groups, mixed and randomly selected to initiate the test.
Condiții experimentale pentru FSDT (specii de apă dulce)
For the three-spined stickleback, see the description of artificial fertilisation in Appendix 11.
Oryzias latipes
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.
Peștele-zebră (Danio rerio)
The test should continue until sexual differentiation in the control group is completed (60 dph for Japanese medaka, the three-spined stickleback and zebrafish).
Ghidrinul (Gasterostreus aculeatus)
Loading
Flux continuu sau semistatic
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).
Becuri fluorescente (spectru larg)
The eggs should be randomly distributed (by using statistical tables for randomisation) among treatments.
Intensitatea luminii
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.
10-20 μE/m2/s, 540-1080 lux sau 50-100 ft-c (niveluri ambientale din laborator)
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.
12-16 h de lumină, 8-12 h de întuneric
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.
Dimensiunea minimă a camerelor de testare
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.
Camerele individuale trebuie să conțină un volum de apă de minimum 7 l
The photoperiod and water temperature should be appropriate for the test species (see Appendix 2 for experimental conditions for the FSDT).
Vârsta organismelor testate la începutul expunerii
Food and feeding are critical, and it is essential that the correct food for each stage is supplied at appropriate time intervals and at a level sufficient to support normal growth.
Ouă recent fertilizate (stadiul de blastulă timpurie)
Feeding should be ad libitum whilst minimising the surplus.
Ouă recent fertilizate
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.
Numărul de ouă per tratament
Surplus food and faeces should be removed, as necessary, to avoid accumulation of waste.
Minimum 3 (plus controale corespunzătoare)
Feeding should be withheld 24 hours before ending the test.
Artemia vii, creveți adulți congelați, hrană sub formă de fulgi etc. Se recomandă administrarea de hrană de două ori pe zi.
Examples of appropriate food items are listed in Appendix 2 (see also the OECD Fish Testing Framework (39).
Hrană specială pentru pești tineri, Artemia, creveți adulți congelați, hrană sub formă de fulgi etc. Se recomandă administrarea de hrană de două ori pe zi.
Test chemicals should be spaced as described in Appendix 4.
Fără aerare, cu excepția cazului în care concentrația OD scade sub 60 % din saturație