natgillin/translations-raw huggingface dataset:
The assay is not designed to identify specific mechanisms of hormonal disruption because the test animals possess an intact hypothalamic-pituitary-gonadal (HPG) axis, which may respond to chemicals that impact on the HPG axis at different levels.
charakteristika rozpúšťadla (povaha, použitá koncentrácia),
The Fish Short Term Reproduction assay (OECD TG 229) includes fecundity and, as appropriate, gonadal histopathology for the fathead minnow, as well as all endpoints included in this test method.
odôvodnenie výberu rozpúšťadla (ak je iné ako voda).
OECD TG 229 provides a screening of chemicals which affect reproduction through various mechanisms including endocrine modalities.
Testovacie živočíchy:
This should be considered prior to selecting the most appropriate test method.
druh a kmeň,
these endpoints are vitellogenin and secondary sexual characteristics.
údaje o postupe aklimatizácie živočíchov,
Vitellogenin is measured in fathead minnow, Japanese medaka and zebrafish, whereas secondary sex characteristics are measured in fathead minnow and Japanese medaka only.
telesná hmotnosť rýb na začiatku expozície (z podvzorky populácie rýb).
This bioassay serves as an in vivo screening assay for certain endocrine modes of action and its application should be seen in the context of the “OECD Conceptual Framework for the Testing and Assessment of Endocrine Disrupting Chemicals” (28).
Podmienky testu:
INITIAL CONSIDERATIONS AND LIMITATIONS
použitý postup testovania (druh testu, stupeň zaťaženia, chovná hustota atď.),
Vitellogenin is normally produced by the liver of female oviparous vertebrates in response to circulating endogenous oestrogen.
metóda prípravy zásobných roztokov a prietok,
It is a precursor of egg yolk proteins and, once produced in the liver, travels in the bloodstream to the ovary, where it is taken up and modified by developing eggs.
nominálne testovacie koncentrácie, týždenne merané koncentrácie testovacích roztokov a použitá analytická metóda, stredné namerané hodnoty a štandardné odchýlky v testovacích nádobách a dôkaz o tom, že merania zodpovedajú koncentráciám testovanej chemikálie v skutočnom roztoku,
Vitellogenin is almost undetectable in the plasma of immature female and male fish because they lack sufficient circulating oestrogen;
charakteristiky riediacej vody (vrátane pH, tvrdosti, alkality, teploty, koncentrácie rozpusteného kyslíka, úrovne reziduálneho chlóru, celkového organického uhlíka, suspendovaných tuhých látok a výsledkov akýchkoľvek iných vykonaných meraní),
however, the liver is capable of synthesizing and secreting vitellogenin in response to exogenous oestrogen stimulation.
kvalita vody v testovacích nádobách:
The measurement of vitellogenin serves for the detection of chemicals with various oestrogenic modes of action.
pH, tvrdosť, teplota a koncentrácia rozpustného kyslíka,
The detection of oestrogenic chemicals is possible via the measurement of vitellogenin induction in male fish, and it has been abundantly documented in the scientific peer-reviewed literature (e.g. (7)).
podrobné informácie o kŕmení [napr. druh potravy, zdroj, dávkované množstvo a frekvencia a analýza príslušných kontaminantov (napr. polychlórované bifenyly, polycyklické aromatické uhľovodíky a organochlórové pesticídy), ak je k dispozícii].
Vitellogenin induction has also been demonstrated following exposure to aromatizable androgens (8, 9).
dôkaz, že kontrolné vzorky splnili kritériá prijateľnosti testu,
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-estradiol, causes a decrease in the vitellogenin level, which is used to detect chemicals having aromatase inhibiting properties (10, 11).
údaje o úmrtnosti vyskytujúcej sa pri každej z testovacích koncentrácií a v kontrolnej vzorke,
The biological relevance of the vitellogenin response following oestrogenic/aromatase inhibition is established and has been broadly documented.
používané štatistické analytické techniky, spracovanie údajov a odôvodnenie použitia daných techník,
However, it is possible that production of VTG in females can also be affected by general toxicity and non-endocrine toxic modes of action, e.g. hepatotoxicity.
údaje o biologických pozorovaniach celkovej morfológie vrátane sekundárnych pohlavných znakov a vitelogenínu,
Several measurement methods have been successfully developed and standardised for routine use.
výsledky analýz údajov, najlepšie v tabuľkovej a grafickej forme,
This is the case of species-specific Enzyme-Linked Immunosorbent Assay (ELISA) methods using immunochemistry for the quantification of vitellogenin produced in small blood or liver samples collected from individual fish (12, 13, 14, 15, 16, 17, 18).
výskyt akýchkoľvek neobvyklých reakcií rýb a akýchkoľvek viditeľných účinkov vyvolaných testovanou chemikáliou.
Fathead minnow blood, zebrafish blood or head/tail homogenate, and medaka liver are sampled for VTG measurement.
USMERNENIE PRE INTERPRETÁCIU A AKCEPTÁCIU VÝSLEDKOV TESTU
In medaka, there is a good correlation between VTG measured from blood and from liver (19).
Táto časť obsahuje niekoľko aspektov, ktoré treba brať do úvahy pri interpretácii výsledkov testu pre jednotlivé merané parametre.
Appendix 6 provides the recommended procedures for sample collection for vitellogenin analysis.
Výsledky by sa mali interpretovať opatrne, ak sa zdá, že testovaná chemikália má zjavné toxické účinky alebo ovplyvňuje celkový stav testovacieho živočícha.
Kits for the measurement of vitellogenin are widely available;
S cieľom umožniť zmysluplnú interpretáciu údajov je potrebné pri stanovení rozsahu testovacích koncentrácií dbať na to, aby sa neprekročila maximálna tolerovaná koncentrácia.
such kits should be based on a validated species-specific ELISA method.
Je dôležité mať najmenej jednu vzorku s aplikovanou chemikáliou, v ktorej nie sú žiadne príznaky toxických účinkov.
Secondary sex characteristics in male fish of certain species are externally visible, quantifiable and responsive to circulating levels of endogenous androgens;
Príznaky choroby a príznaky toxických účinkov by sa mali dôkladne vyhodnotiť a zaznamenať.
this is the case for the fathead minnow and the medaka — but not for zebrafish, which does not possess quantifiable secondary sex characteristics.
Je napríklad možné, že produkciu vitelogenínu v telách samičiek môže ovplyvňovať aj všeobecná toxicita a neendokrinné toxické mechanizmy pôsobenia, napríklad hepatotoxicita.
Females maintain the capacity to develop male secondary sex characteristics, when they are exposed to androgenic chemicals in water.
Interpretáciu účinkov však môžu podporiť iné množstvá aplikovanej chemikálie, ktoré nie je možné si pomýliť so systémovou toxicitou.
Several studies are available in the scientific literature to document this type of response in fathead minnow (20) and medaka (21).
Existuje niekoľko aspektov, ktoré treba brať do úvahy pri akceptácii výsledkov testu.
A decrease in secondary sex characteristics in males should be interpreted with caution because of low statistical power, and should be based on expert judgement and weight of evidence.
Ako usmernenie platí, že úrovne vitelogenínu v kontrolných skupinách samčekov a samičiek by mali byť odlišné a líšiť by sa mali približne o tri rádové hodnoty v prípade čereble potočnej a dánia pruhovaného a približne o jeden rád v prípade medaky.
There are limitations to the use of zebrafish in this assay, due to the absence of quantifiable secondary sex characteristics responsive to androgenic acting chemicals.
Príklady rozsahu hodnôt, ktoré sa vyskytli v kontrolných skupinách a v skupinách s aplikovanou chemikáliou, sú k dispozícii vo validačných správach (1), (2), (3), (4).
In the fathead minnow, the main indicator of exogenous androgenic exposure is the number of nuptial tubercles located on the snout of the female fish.
Vysoké hodnoty vitelogenínu pri samčekoch v kontrolných skupinách by mohli ohroziť citlivosť testu a jeho schopnosť odhaliť slabé agonisty estrogénu.
In the medaka, the number of papillary processes constitutes the main marker of exogenous exposure to androgenic chemicals in female fish.
Nízke hodnoty vitelogenínu pri samičkách v kontrolných skupinách by mohli ohroziť citlivosť testu a jeho schopnosť odhaliť inhibítory aromatázy a antagonisty estrogénu.
Appendix 5A and Appendix 5B indicate the recommended procedures to follow for the evaluation of sex characteristics in fathead minnow and in medaka, respectively.
Na vypracovanie tohto usmernenia sa použili validačné štúdie.
Definitions used in this test method are given in Appendix 1.
Ak laboratórium predtým nevykonávalo tento test alebo nastali podstatné zmeny (napr. zmena kmeňa rýb alebo dodávateľa), je vhodné realizovať štúdiu technickej spôsobilosti.
In the assay, male and female fish in a reproductive status are exposed together in test vessels.
Odporúča sa použitie chemikálií, ktoré zahŕňajú rôzne mechanizmy pôsobenia alebo vplyvu na viaceré parametre testu.
Their adult and reproductive status enables a clear differentiation of each sex, and thus a sex-related analysis of each endpoint, and ensures their sensitivity towards exogenous chemicals.
V praxi sa každému laboratóriu odporúča, aby si budovalo svoju vlastnú databázu historických kontrolných údajov pre samčekov a samičky a aby používalo pozitívnu kontrolnú chemikáliu na estrogénne pôsobenie (napr. 17β-estradiol, 100 ng/l, alebo známe slabé agonisty), ktoré vedie k zvýšeniu množstva vitelogenínu v telách samčekov rýb, pozitívnu kontrolnú chemikáliu na inhibíciu aromatázy (napr. fadrozol alebo prochloraz, 300 μg/l), ktorá vedie k zníženiu množstva vitelogenínu v telách samičiek rýb, a pozitívnu kontrolnú chemikáliu na androgénne pôsobenie (napr. 17β-trenbolón, 5 μg/l), ktoré vedie k vzniku sekundárnych pohlavných znakov pri samičkách čereble potočnej a medaky.
At test termination, sex is confirmed by macroscopic examination of the gonads following ventral opening of the abdomen with scissors.
Všetky tieto údaje je možné porovnať s dostupnými údajmi z validačných štúdií (1), (2), (3), aby sa zabezpečila odbornosť laboratória.
An overview of the relevant bioassay conditions is provided in Appendix 2.
Vo všeobecnosti by sa malo meranie vitelogenínu považovať za pozitívne, ak existuje štatisticky významné zvýšenie množstva vitelogenínu v telách samčekov (p < 0,05) alebo štatisticky významný pokles v telách samičiek (p < 0,05) v porovnaní s kontrolnou skupinou, aspoň pri najvyššej testovacej dávke, a ak sa nevyskytnú príznaky všeobecnej toxicity.
The assay is normally initiated with fish sampled from a population that is in spawning condition;
Pozitívny výsledok ďalej podporí preukázanie biologicky hodnoverného vzťahu medzi dávkou a reakciou.
senescent animals should not be used.
Ako už bolo uvedené, pokles množstva vitelogenínu nemusí mať výlučne endokrinný pôvod.
Guidance on the age of fish and on the reproductive status is provided in the section on Selection of fish.
Pozitívny výsledok by sa však mal vo všeobecnosti interpretovať ako dôkaz endokrinného pôsobenia in vivo a mal by spravidla viesť k opatreniam na ďalšie objasnenie.
The assay is conducted using three chemical exposure concentrations as well as a water control, and a solvent control if necessary.
Vitellogenin induction by 17ß-estradiol and 17ß-ethinylestradiol in male zebrafish (Danio rerio).
Two vessels or replicates per treatment are used (each vessel containing 5 males and 5 females) in medaka and zebrafish, whereas four vessels or replicates per treatment are used (each vessel containing 2 males and 4 females) in fathead minnow.
Screening and testing for endocrine disruption in fish-biomarkers as ‚signposts,‘ not ‚traffic lights,‘ in risk assessment.
This is to accommodate the territorial behaviour of male fathead minnow while maintaining sufficient power of the assay.
Villeneuve a G.T.
The exposure is conducted for 21 days and sampling of fish is performed at day 21 of exposure.
Skratky a vymedzenie pojmov
On sampling at day 21, all animals are killed humanely.
Chemikália
Secondary sex characteristics are measured in fathead minnow and medaka (see Appendix 5A and Appendix 5B);
látka alebo zmes.
blood samples are collected for determination of vitellogenin in zebrafish and fathead minnow, alternatively head/tail can be collected for the determination of vitellogenin in zebrafish (Appendix 6);
variačný koeficient.
liver is collected for VTG analysis in medaka (Appendix 6).
enzýmové imunosorbentové stanovenie.
TEST ACCEPTANCE CRITERIA
Stupeň zaťaženia
For the test results to be acceptable the following conditions apply:
čerstvá hmotnosť rýb na objem vody.
the mortality in the water (or solvent) controls should not exceed 10 % at the end of the exposure period;
Chovná hustota
the dissolved oxygen concentration should be at least 60 % of the air saturation value (ASV) throughout the exposure period;
počet rýb na objem vody.
the water temperature should not differ by more than ± 1,5 °C between test vessels at any one time during the exposure period and be maintained within a range of 2 °C within the temperature ranges specified for the test species (Appendix 2);
Vitelogenín (VTG)
evidence should be available to demonstrate that the concentrations of the test chemical in solution have been satisfactorily maintained within ± 20 % of the mean measured values.
fosfolipoglykoproteín, slúžiaci ako prekurzor proteínov vaječného žĺtka, ktorý sa normálne vyskytuje v telách pohlavne aktívnych samičiek všetkých vajcorodých druhov.
Normal laboratory equipment and especially the following:
Os HPG
oxygen and pH meters;
os hypothalamus – hypofýza – gonády (hypothalamic-pituitary-gonadal).
equipment for determination of water hardness and alkalinity;
MTC (maximum tolerated concentration)
adequate apparatus for temperature control and preferably continuous monitoring;
maximálna tolerovaná koncentrácia, ktorá predstavuje približne 10 % hodnoty LC50.
tanks made of chemically inert material and of a suitable capacity in relation to the recommended loading and stocking density (see Appendix 2);
Experimentálne podmienky skríningového testu na endokrinnom systéme rýb
spawning substrate for fathead minnow and zebrafish, Appendix 4 gives the necessary details;
Odporúčané druhy
suitably accurate balance (i.e. accurate to ± 0,5 mg).
Čerebľa potočná
Any water in which the test species shows suitable long-term survival and growth may be used as test water.
Dánio pruhované
It should be of constant quality during the period of the test.
Druh testu
The pH of the water should be within the range 6,5 to 8,5, but during a given test it should be within a range of ± 0,5 pH units.
prietokový
In order to ensure that the dilution water will not unduly influence the test result (for example by complexion of test chemical), samples should be taken at intervals for analysis.
Teplota vody
Measurements of heavy metals (e.g. Cu, Pb, Zn, Hg, Cd, and Ni), major anions and cations (e.g. Ca2+, Mg2+, Na+, K+, Cl-, and SO42-), pesticides (e.g. total organophosphorus and total organochlorine pesticides), total organic carbon and suspended solids should be made, for example, every three months where dilution water is known to be relatively constant in quality.
Kvalita osvetlenia
If water quality has been demonstrated to be constant over at least one year, determinations can be less frequent and intervals extended (e.g. every six months).
Žiarivky (široké spektrum)
Some chemical characteristics of acceptable dilution water are listed in Appendix 3.
10 – 20 μE/m2/s, 540 – 1000 luxov alebo 50 – 100 ft-c (úrovne v laboratórnom prostredí)
Test solutions of the chosen concentrations are prepared by dilution of a stock solution.
Fotoperióda (možné sú prechody svitania/súmraku, ale nepovažujú sa za nutné)
The stock solution should preferably be prepared by simply mixing or agitating the test chemical in dilution water by using mechanical means (e.g. stirring or ultrasonication).
16 hod. svetlo, 8 hod. tma
Saturation columns (solubility columns) can be used for achieving a suitable concentrated stock solution.
12 – 16 hodín svetla a 12 – 8 hodín tmy
The use of a solvent carrier is not recommended.
< 5 g/l
However, in case a solvent is necessary, a solvent control should be run in parallel, at the same solvent concentration as the chemical treatments.
Veľkosť skúšobnej komory
For difficult test chemicals, a solvent may be technically the best solution;
10 l (minimálne)
the OECD Guidance Document on aquatic toxicity testing of difficult substances and mixtures should be consulted (22).
2 l (minimálne)
The choice of solvent will be determined by the chemical properties of the chemical.
5 l (minimálne)
The OECD Guidance Document recommends a maximum of 100 μl/l, which should be observed.
Objem testovacieho roztoku
However a recent review (23) highlighted additional concerns when using solvents for endocrine activity testing.
8 l (minimálne)
Therefore it is recommended that the solvent concentration, if necessary, is minimised wherever technically feasible (dependent on the physical-chemical properties of the test chemical).
1,5 l (minimálne)
A flow-through test system will be used.
4 l (minimálne)
Such a system continually dispenses and dilutes a stock solution of the test chemical (e.g. metering pump, proportional diluter, saturator system) in order to deliver a series of concentrations to the test chambers.
Výmena objemu testovacích roztokov
The flow rates of stock solutions and dilution water should be checked at intervals, preferably daily, during the test and should not vary by more than 10 % throughout the test.
minimálne 6 denne
Care should be taken to avoid the use of low-grade plastic tubing or other materials that may contain biologically active chemicals.
Minimálne päťkrát denne
When selecting the material for the flow-through system, possible adsorption of the test chemical to this material should be considered.
pozri bod 20
Holding of fish
samčekovia:
Test fish should be selected from a laboratory population, preferably from a single stock, which has been acclimated for at least two weeks prior to the test under conditions of water quality and illumination similar to those used in the test.
samičky:
It is important that the loading rate and stocking density (for definitions, see Appendix 1) be appropriate for the test species used (see Appendix 2).
Počet rýb v každej testovacej nádobe
Following a 48-hour settling-in period, mortalities are recorded and the following criteria applied:
6 (2 samčekovia a 4 samičky)
mortalities of greater than 10 % of population in seven days:
10 (5 samčekov a 5 samičiek)
reject the entire batch;
Počet testovaných koncentrácií
mortalities of between 5 % and 10 % of population:
= 3 (plus primerané kontrolné vzorky)
acclimation for seven additional days;
Počet nádob na každú aplikáciu chemikálie
if more than 5 % mortality during second seven days, reject the entire batch;
minimálne 4
mortalities of less than 5 % of population in seven days:
minimálne 2
accept the batch
Počet rýb na každú testovanú koncentráciu
A one-week pre-exposure period is recommended, with animals placed in vessels similar to the actual test.
Kŕmny režim
Fish should be fed ad libitum throughout the holding period and during the exposure phase.
živé alebo mrazené dospelé jedince alebo larvy žiabronôžky soľnej, dva- alebo trikrát denne (ad libitum), komerčne dostupná potrava, alebo kombinácia uvedeného
The exposure phase is started with sexually dimorphic adult fish from a laboratory supply of reproductively mature animals (e.g. with clear secondary sexual characteristics visible as far as fathead minnow and medaka are concerned), and actively spawning.
larvy žiabronôžky soľnej, dva- alebo trikrát denne (ad libitum), komerčne dostupná potrava, alebo kombinácia uvedeného
For general guidance only (and not to be considered in isolation from observing the actual reproductive status of a given batch of fish), fathead minnows should be approximately 20 (± 2) weeks of age, assuming they have been cultured at 25 ± 2 °C throughout their lifespan.
žiadne, pokiaľ koncentrácia rozpusteného kyslíka neklesne pod 60 % hodnoty saturácie vzduchom
Japanese medaka should be approximately 16 (± 2) weeks of age, assuming they have been cultured at 25 ± 2 °C throughout their lifespan.
s čistým povrchom, studňová alebo rekonštituovaná voda alebo odchlórovaná voda z vodovodu
Zebrafish should be approximately 16 (± 2) weeks of age, assuming they have been cultured at 26 ± 2 °C throughout their lifespan.
Obdobie pred expozíciou
TEST DESIGN
odporúča sa 7 dní
Three concentrations of the test chemical, one control (water) and, if needed, one solvent control are used.
Trvanie expozície chemikálii
The data may be analysed in order to determine statistically significant differences between treatment and control responses.
Biologické sledované parametre
These analyses will inform whether further longer term testing for adverse effects (namely, survival, development, growth and reproduction) is required for the chemical, rather than for use in risk assessment (24).
prežitie
For zebrafish and medaka, on day 21 of the experiment, males and females from each treatment level (5 males and 5 females in each of the two replicates) and from the control(s) are sampled for the measurement of vitellogenin and secondary sex characteristics, where applicable.
správanie
For fathead minnow, on day 21 of exposure, males and females (2 males and 4 females in each of the four replicates) and from the control(s) are sampled for the measurement of vitellogenin and secondary sex characteristics.
sekundárne pohlavné znaky
For the purposes of this test, the highest test concentration should be set by the maximum tolerated concentration (MTC) determined from a range finder or from other toxicity data, or 10 mg/l, or the maximum solubility in water, whichever is lowest.
vitelogenín
The MTC is defined as the highest test concentration of the chemical which results in less than 10 % mortality.
Prijateľnosť testu
Using this approach assumes that there are existing empirical acute toxicity data or other toxicity data from which the MTC can be estimated.
rozpustený kyslík > 60 % saturácie;
Estimating the MTC can be inexact and typically requires some professional judgment.
stredná teplota 25 °C ± 2 °C;
Three test concentrations, spaced by a constant factor not exceeding 10, and a dilution-water control (and solvent control if necessary) are required.
prežitie rýb v kontrolných vzorkách 90 %;
A range of spacing factors between 3,2 and 10 is recommended.
merané testovacie koncentrácie v rozsahu 20 % stredných meraných hodnôt pre každú úroveň aplikácie chemikálie
Selection and weighing of test fish
stredná teplota 24 °C ± 2 °C;
It is important to minimise variation in weight of the fish at the beginning of the assay.
stredná teplota 26 °C ± 2 °C;
Suitable size ranges for the different species recommended for use in this test are given in Appendix 2.
Niektoré chemické vlastnosti prijateľnej riediacej vody
For the whole batch of fish used in the test, the range in individual weights for male and female fish at the start of the test should be kept, if possible, within ± 20 % of the arithmetic mean weight of the same sex.
< 20 mg/l
It is recommended to weigh a subsample of the fish stock before the test in order to estimate the mean weight.
< 2 mg/l
Conditions of exposure
Celkové organofosfátové pesticídy
The test duration is 21 days, following a pre-exposure period.
Dodatok 4A
The recommended pre-exposure period is one week.
Substrát na neresenie pre dánio pruhované
Fish should be fed ad libitum with an appropriate food (Appendix 2) at a sufficient rate to maintain body condition.
Nádoba na neresenie
Care should be taken to avoid microbial growth and water turbidity.
celosklená nádoba s rozmermi napríklad 22 × 15 × 5,5 cm (dĺžka × šírka × výška) pokrytá odnímateľnou drôtenou mriežkou z nehrdzavejúcej ocele (so šírkou otvorov 2 mm).
As a general guidance, the daily ration may be divided into two or three equal portions for multiple feeds per day, separated by at least three hours between each feed.
Mriežka by mala pokrývať otvor nádoby v rovine položenej nižšie, ako je okraj nádoby.