natgillin/translations-raw huggingface dataset:
The biological relevance of the vitellogenin response following oestrogenic/aromatase inhibition is established and has been broadly documented.
výskyt akýchkoľvek neobvyklých reakcií rýb a akýchkoľvek viditeľných účinkov vyvolaných testovanou chemikáliou.
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.
USMERNENIE KINTERPRETÁCII A AKCEPTÁCII VÝSLEDKOV TESTU
Several measurement methods have been successfully developed and standardised for routine use.
Táto časť obsahuje niekoľko aspektov, ktoré treba brať do úvahy pri interpretácii výsledkov testu pre jednotlivé merané ukazovatele.
This is the case of species-specific Enzyme-Linked Immunosorbent Assay (ELISA) methods using immunochemistry for the quantification of VTG produced in small blood or liver samples collected from individual fish (12, 13, 14, 15, 16, 17, 18).
Výsledky by sa mali interpretovať opatrne, ak sa zdá, že testovaná chemikália má zjavné toxické účinky alebo ovplyvňuje celkový stav testovaného zvieraťa.
Fathead minnow blood, zebrafish blood or head/tail homogenate, and medaka liver are sampled for VTG measurement.
S cieľom umožniť zmysluplnú interpretáciu údajov by sa pri stanovení rozsahu testovacích koncentrácií malo dbať na to, aby sa neprekročila maximálna tolerovaná koncentrácia.
In medaka, there is a good correlation between VTG measured from blood and from liver (19).
Je dôležité mať najmenej jednu skupinu s aplikovanou chemikáliou, v ktorej nie sú žiadne príznaky toxických účinkov.
Appendix 6 provides the recommended procedures for sample collection for VTG analysis.
Príznaky choroby a príznaky toxických účinkov by sa mali dôkladne vyhodnotiť a zaznamenať.
Kits for the measurement of VTG are widely available; such kits should be based on a validated species-specific ELISA method.
Je napríklad možné, že produkciu VTG v telách samíc môže ovplyvňovať aj všeobecná toxicita a neendokrinné toxické mechanizmy pôsobenia, napríklad hepatotoxicita.
Secondary sex characteristics in male fish of certain species are externally visible, quantifiable and responsive to circulating levels of endogenous androgens; this is the case for the fathead minnow and the medaka — but not for zebrafish which does not possess quantifiable secondary sex characteristics.
Interpretáciu účinkov však možno podporiť inými množstvami aplikovanej chemikálie, ktoré si nemožno pomýliť so systémovou toxicitou.
Females maintain the capacity to develop male secondary sex characteristics, when they are exposed to androgenic chemicals in water.
Existuje niekoľko aspektov, ktoré treba brať do úvahy pri akceptácii výsledkov testu.
Several studies are available in the scientific literature to document this type of response in fathead minnow (20) and medaka (21).
Ako usmernenie platí, že úrovne VTG v kontrolných skupinách samcov a samíc by sa mali odlišovať a mali by ich oddeľovať približne tri rádové hodnoty v prípade čereble potočnej a dania pruhovaného a približne jedna rádová hodnota v prípade ryžovníčky japonskej.
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.
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).
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.
Vysoké hodnoty VTG kontrolných samcov by mohli ohroziť citlivosť testu a jeho schopnosť odhaliť slabé agonisty estrogénu.
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.
Nízke hodnoty VTG samíc v kontrolných skupinách by mohli ohroziť citlivosť testu a jeho schopnosť odhaliť inhibítory aromatázy a antagonisty estrogénu.
In the medaka, the number of papillary processes constitutes the main marker of exogenous exposure to androgenic chemicals in female fish.
Na vypracovanie tohto usmernenia sa použili validačné štúdie.
Appendix 5A and Appendix 5B indicate the recommended procedures to follow for the evaluation of sex characteristics in fathead minnow and in medaka, respectively.
Kvantifikácia tvorby vajíčok podlieha značnej variabilite [variačný koeficient (CV) sa môže pohybovať od 20 do 60 %], ktorá ovplyvňuje schopnosť testu odhaliť významný pokles produkcie vajíčok menší ako 70 % v prípade, že sa CV blíži k 50 % alebo je vyšší.
The 21-day fish assay includes the evaluation of quantitative egg production and preservation of gonads for optional histopathology examination.
Keď je CV obmedzený na nižšie hodnoty (okolo 20 – 30 %), potom bude mať test prijateľnú silu (80 %) na odhalenie 40 – 50 % poklesu tvorby vajíčok.
Some regulatory authorities may require this additional endpoint for a more complete evaluation of the reproductive fitness of the test animals, or in cases where vitellogenin and secondary sex characteristics did not respond to the chemical exposure.
Plán štúdie použitý pre čerebľu potočnú, zahŕňajúci štyri replikáty na úroveň aplikácie chemikálie, by mal umožniť väčšiu silu vo vzťahu k ukazovateľu plodivosti v porovnaní s plánom testu len s 2 replikátmi.
Although some endpoints may be highly diagnostic (e.g. VTG induction in males and tubercle formation in females), not all endpoints (e.g. fecundity and gonad histopathology) in the assay are intended to unequivocally identify specific cellular mechanisms of action.
Ak laboratórium predtým nevykonávalo tento test alebo nastali podstatné zmeny (napr. zmena kmeňa rýb alebo dodávateľa), je vhodné realizovať štúdiu technickej spôsobilosti.
Rather, the suite of endpoints, collectively, allows inferences to be made with regard to possible endocrine disturbances and thus provide guidance for further testing.
Odporúča sa použitie chemikálií, ktoré zahŕňajú rôzne mechanizmy pôsobenia alebo vplyvu na viaceré ukazovatele testu.
Although not endocrine specific, fecundity, due to its demonstrated sensitivity across known endocrine active chemicals (5), is an important endpoint to include because when it and other endpoints are unaffected one is more confident that a compound is not likely endocrine active.
V praxi sa každému laboratóriu odporúča, aby si budovalo svoju vlastnú historickú databázu kontrolných údajov pre samcov a samice a aby používalo chemikáliu na pozitívnu kontrolu na estrogénne pôsobenie (napr. 17β-estradiol, 100 ng/l, alebo známe slabé agonisty), ktoré vedie k zvýšeniu množstva VTG u samcov rýb, chemikáliu na pozitívnu kontrolu na inhibíciu aromatázy (napr. fadrozol alebo prochloraz, 300 μg/l), ktorá vedie k zníženiu množstva VTG u samíc rýb, a chemikáliu na pozitívnu kontrolu na androgénne pôsobenie (napr. 17β-trenbolón, 5 μg/l), ktoré vedie k vzniku sekundárnych pohlavných znakov u samíc čereble potočnej a ryžovníčky japonskej.
However, when fecundity is affected it will contribute heavily in weight of evidence inferences.
Všetky tieto údaje možno porovnať s dostupnými údajmi z validačných štúdií (1, 2, 3), aby sa zabezpečila spôsobilosť laboratória.
Guidance on data interpretation and acceptance of test results is provided further in this test method.
Vo všeobecnosti by sa meranie VTG malo považovať za pozitívne, ak v porovnaní s kontrolnou skupinou existuje štatisticky významné zvýšenie množstva VTG u samcov (p < 0,05) alebo štatisticky významný pokles u samíc (p < 0,05) aspoň pri najvyššej testovanej dávke a ak sa nevyskytnú príznaky všeobecnej toxicity.
Definitions used in this test method are given in Appendix 1.
Pozitívny výsledok ďalej podporí preukázanie biologicky hodnoverného vzťahu medzi dávkou a reakčnou krivkou.
In the assay, male and female fish in a reproductive status are exposed together in test vessels.
Ako už bolo uvedené, pokles VTG nemusí mať výlučne endokrinný pôvod; 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.
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.
Regulačné orgány môžu vyžadovať vyhodnotenie histopatológie gonád s cieľom stanoviť reprodukčné zdravie testovaných zvierat a umožniť posúdenie váhy dôkazov vyplývajúcich z výsledkov testu.
At test termination, sex is confirmed by macroscopic examination of the gonads following ventral opening of the abdomen with scissors.
Výsledky histopatológie gonád nemusia byť nevyhnutné v prípadoch, keď buď VTG alebo sekundárne pohlavné znaky sú pozitívne (t. j. zvýšenie alebo pokles VTG alebo vznik sekundárnych pohlavných znakov).
An overview of the relevant bioassay conditions are provided in Appendix 2.
OECD (2006b).
The assay is normally initiated with fish sampled from a population that is in spawning condition; senescent animals should not be used.
OECD (2007).
Guidance on the age of fish and on the reproductive status is provided in the section on Selection of fish.
Owens JW (2007).
The assay is conducted using three chemical exposure concentrations as well as a water control, and a solvent control if necessary.
US EPA (2007).
Two vessels or replicates per treatment are used for zebrafish (each vessel containing 5 males and 5 females).
OECD (2008).
Four vessels or replicates per treatment are used for fathead minnow (each vessel containing 2 males and 4 females).
Sumpter J.P. a S. Jobling (1995).
This is to accommodate the territorial behaviour of male fathead minnow while maintaining sufficient power of the assay.
Pawlowski S., et al. (2004).
Four vessels or replicates per treatment are used for medaka (each vessel containing 3 males and 3 females).
Andersen L., et al. (2006).
The exposure is conducted for 21-days and sampling of fish is performed at day 21 of exposure.
Ankley G.T., et al. (2002).
Quantitative fecundity is monitored daily.
Panter G.H., et al. (2004).
On sampling at day 21, all animals are killed humanely.
Parks L.G., et al. (1999).
Secondary sex characteristics are measured in fathead minnow and medaka (see Appendix 5A and Appendix 5B); blood samples are collected for determination of VTG in zebrafish and fathead minnow, alternatively head/tail can be collected for the determination of VTG in zebrafish (Appendix 6); liver is collected for VTG analysis in medaka (Appendix 6); gonads are fixed either in whole or dissected for potential histopathological evaluation (22).
Part C Pharmacology, toxicology and endocrinology; 123(2): 113 – 25.
TEST ACCEPTANCE CRITERIA
Fenske M., et al. (2001).
For the test results to be acceptable the following conditions apply:
C 129 (3): 217 – 232.
the mortality in the water (or solvent) controls should not exceed 10 per cent at the end of the exposure period;
Holbech H., et al. (2001).
the dissolved oxygen concentration should be at least 60 per cent of the air saturation value (ASV) throughout the exposure period;
Vitellogenin induction by 17β-estradiol and 17α-ethinylestradiol in male zebrafish (Danio rerio).
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);
C 131: 531 – 539.
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;
Brion F., et al. (2002).
evidence that fish are actively spawning in all replicates prior to initiating chemical exposure and in control replicates during the test.
Development and validation of an enzyme-linked immunosorbent assay to measure vitellogenin in the zebrafish (Danio rerio).
oxygen and pH meters;
Yokota H., et al. (2001).
equipment for determination of water hardness and alkalinity;
Development of an ELISA for determination of the hepatic vitellogenin in Medaka (Oryzias latipes).
adequate apparatus for temperature control and preferably continuous monitoring;
Effects of the androgenic growth promoter 17-beta-trenbolone on fecundity and reproductive endocrinology of the fathead minnow.
tanks made of chemically inert material and of a suitable capacity in relation to the recommended loading and stocking density (see Appendix 2);
Environmental Toxicology and Chemistry; 22(6): 1350 – 60.
spawning substrate for fathead minnow and zebrafish, Appendix 4 gives the necessary details.
OECD (2010).
suitably accurate balance (i.e. accurate to ± 0,5 mg).
Guidance Document on Fish Gonadal Histopathology.
Any water in which the test species shows suitable long-term survival and growth may be used as test water.
OECD Environmental Health and Safety Publications Series on Testing and Assessment No.
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.
123, Paris.
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.
OECD (2000) Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures.
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.
Series on Testing and Assessment No.
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).
Paris
Some chemical characteristics of acceptable dilution water are listed in Appendix 3.
Miles-Richardson S.R., et al. (1999).
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).
47, 129 – 145.
Saturation columns (solubility columns) can be used for achieving a suitable concentrated stock solution.
Martinovic D., et al. (2008).
However, in case a solvent is necessary, a solvent control should be run in parallel, at the same solvent concentration as the chemical treatments.
Chem.
For difficult to test chemicals, a solvent may be technically the best solution; the OECD guidance document on aquatic toxicity testing of difficult substances and mixtures should be consulted (23).
27, 478-488.
The choice of solvent will be determined by the chemical properties of the substance or mixture.
OECD (2012), OECD Conceptual Framework for Testing and Assessment of Endocrine Disrupters, OECD Environmental Health and Safety Publications, Series on Testing and Assessment No.
The OECD guidance document recommends a maximum of 100 μl/l, which should be observed.
150, OECD, Paris.
However a recent review (24) highlighted additional concerns when using solvents for endocrine activity testing.
SKRATKY A DEFINÍCIE
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).
látka alebo zmes
A flow-through test system will be used.
variačný koeficient
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.
enzýmové imunosorbentové stanovenie
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.
Veľkosť násady
Care should be taken to avoid the use of low-grade plastic tubing or other materials that may contain biologically active chemicals.
Veľkosť násady mokrá hmotnosť rýb na objem vody
When selecting the material for the flow-through system, possible adsorption of the test chemical to this material should be considered.
maximálna tolerovaná koncentrácia, ktorá predstavuje približne 10 % hodnoty LC50
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.
Chovná hustota
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).
je počet rýb na objem vody
mortalities of greater than 10 % of population in seven days: reject the entire batch;
vitelogenín je fosfolipoglykoproteín, slúžiaci ako prekurzor proteínov vaječného žĺtka, ktorý sa normálne vyskytuje v telách pohlavne aktívnych samíc všetkých vajcorodých druhov.
mortalities of between 5 % and 10 % of population: acclimation for seven additional days; if more than 5 % mortality during second seven days, reject the entire batch;
EXPERIMENTÁLNE PODMIENKY SKRÍNINGU NA ENDOKRINNOM SYSTÉME RÝB
mortalities of less than 5 % of population in seven days: accept the batch.
Typ testu
Fish should not receive treatment for disease during the acclimation period, in the pre-exposure period, or during the exposure period.
prietokový
Pre-exposure and selection of fish
Teplota vody
The one to two-week pre-exposure period is recommended with animals placed in vessels similar to the actual test.
Kvalita osvetlenia
Fish should be fed ad libitum throughout the holding period and during the exposure phase.
Žiarivky (širokospektrálne)
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.
Intenzita svetla
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.
10 – 20 μE/m2/s, 540 – 1000 luxov alebo 50 – 100 ft-c (úrovne v laboratórnom prostredí)
Japanese medaka should be approximately 16 (± 2) weeks of age, assuming they have been cultured at 25 ± 2 °C throughout their lifespan.
12 – 16 hodín svetla a 12 – 8 hodín tmy
Zebrafish should be approximately 16 (± 2) weeks of age, assuming they have been cultured at 26 ± 2 °C throughout their lifespan.
< 5 g/l
Egg production should be assessed daily during the pre-exposure phase.
Veľkosť skúšobnej komory
It is recommended that spawning be observed in all replicate tanks prior to inclusion in the exposure phase of the assay.
10 l (minimálne)
Quantitative guidance on desirable daily egg production cannot be provided at this stage, but it is relatively common to observe average spawns of > 10 eggs/female/day for each species.
2 l (minimálne)
A randomised block design according to egg production output should be used to allocate replicates to the various experimental levels to ensure balanced distribution of replicates.
5 l (minimálne)
TEST DESIGN
Objem testovacieho roztoku
Three concentrations of the test chemical, one control (water) and, if needed, one solvent control are used.
8 l (minimálne)
The data may be analysed in order to determine statistically significant differences between treatment and control responses.
1,5 l (minimálne)
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 (25).
4 l (minimálne)
For zebrafish, 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.
Výmeny objemu testovacích roztokov
For medaka, on day 21 of the experiment, males and females from each treatment level (3 males and 3 females in each of the four replicates) and from the control(s) are sampled for the measurement of vitellogenin and secondary sex characteristics.
minimálne 6 denne
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.
minimálne 5 denne
Quantitative assessment of fecundity is required, and gonadal tissues should be fixed in whole or dissected for potential histopathological evaluation, if required.
Vek testovaných organizmov
Selection of test concentrations
pozri bod 21
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.
Približná mokrá hmotnosť dospelých rýb (g)
The MTC is defined as the highest test concentration of the chemical which results in less than 10 % mortality.
samice: 1,5 ± 20 %
Using this approach assumes that there are existing empirical acute toxicity data or other toxicity data from which the MTC can be estimated.
samce 2,5 ± 20 %
Estimating the MTC can be inexact and typically requires some professional judgment.
samice: 0,35 ± 20 %
Three test concentrations, spaced by a constant factor not exceeding 10, and a dilution-water control (and solvent control if necessary) are required.
samce 0,35 ± 20 %
A range of spacing factors between 3,2 and 10 is recommended.
samice: 0,65 ± 20 %
Selection and weighing of test fish
samce 0,4 ± 20 %
It is important to minimise variation in weight of the fish at the beginning of the assay.
Počet rýb na testovaciu nádobu
Suitable size ranges for the different species recommended for use in this test are given in Appendix 2.
6 (2 samce a 4 samice)
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.
6 (3 samce a 3 samice)
It is recommended to weigh a subsample of the fish stock before the test in order to estimate the mean weight.
10 (5 samci a 5 samíc)
The test duration is 21 days, following a pre-exposure period.
Počet skupín s aplikovanou chemikáliou
The recommended pre-exposure period is one to two weeks.
= 3 (plus primerané kontrolné skupiny)
Fish should be fed ad libitum with an appropriate food (Appendix 2) at a sufficient rate to maintain body condition.
Počet nádob na každú úroveň aplikácie chemikálie
Care should be taken to avoid microbial growth and water turbidity.
najmenej 4
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.
najmenej 2
A single larger ration is acceptable particularly for weekends.
Počet rýb na každú testovaciu koncentráciu
Food should be withheld from the fish for 12 hours prior to sampling/necropsy.
16 dospelých samíc a 8 samcov (4 samice a 2 samci v každej nádobe s replikátom)
Fish food should be evaluated for the presence of contaminants such as organochlorine pesticides, polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs).
12 dospelých samíc a 12 samcov (3 samice a 3 samci v každej nádobe s replikátom)
Food with an elevated level of phytoestrogens that would compromise the response of the assay to known oestrogen agonist (e.g. 17β- estradiol) should be avoided.
10 dospelých samíc a 10 samcov (5 samíc a 5 samci v každej nádobe s replikátom)
Uneaten food and faecal material should be removed from the test vessels at least twice weekly, e.g. by carefully cleaning the bottom of each tank using a siphon.
Kŕmny režim