natgillin/translations-raw huggingface dataset:
Since spawning groups of zebrafish may occasionally fail to spawn, the parallel use of at least three spawning tanks is recommended.
Ocene toksičnosti morajo temeljiti na dolžini glavnega poganjka in treh dodatnih merjenih spremenljivkah (tj. po možnosti skupna dolžina poganjka, sveža teža, suha teža ali število spiralasto razvrščenih listov (glej odstavek 37 in opombe 2, 4 in 5 k temu odstavku)), saj lahko nekatere kemikalije bistveno bolj vplivajo na druge merjene spremenljivke kot na dolžino glavnega poganjka.
To avoid genetic bias, eggs are collected from a minimum of three breeding groups, mixed and randomly selected.
Ta učinek se ne bi zaznal zgolj z izračunom dolžine glavnega poganjka.
To prevent predation of eggs by adult zebrafish, the spawn traps are covered with inert wire mesh of appropriate mesh size (approx.
povprečna specifična hitrost rasti od časa i do časa j,
2 ± 0,5 mm).
merjena spremenljivka v preskusni posodi ali kontrolni posodi ob času i,
If considered necessary, artificial plants made of inert material (e.g., plastic or glass) can be fixed to the mesh as spawning stimulus (3)(4)(5)(23)(35).
merjena spremenljivka v preskusni posodi ali kontrolni posodi ob času j,
Weathered plastic materials which do not leach (e.g., phthalates) should be used.
t časovno obdobje od i do j.
Mating, spawning and fertilisation take place within 30 min after the onset of light and the spawn traps with the collected eggs can be carefully removed.
Za vsako tretirano in kontrolno skupino se izračuna aritmetično povprečje hitrosti rasti in oceni varianca.
Rinsing eggs with reconstituted water after collection from spawning traps is recommended.
Povprečna specifična hitrost rasti se izračuna za celotno preskusno obdobje (čas „i“ v zgornji enačbi pomeni začetek preskusa, čas „j“ pa konec preskusa).
Egg differentiation
Za vsako preskusno koncentracijo in kontrolo se izračuna aritmetično povprečje specifične hitrosti rasti in oceni varianca.
At 26 °C, fertilised eggs undergo the first cleavage after about 15 min and the consecutive synchronous cleavages form 4, 8, 16 and 32 cell blastomers (see Appendix 3)(35).
Oceni se tudi presečna hitrost rasti, da se ocenijo učinki preskusne kemikalije, ki se pojavljajo med obdobjem izpostavljenosti (npr. s pregledovanjem logaritemsko transformiranih krivulj rasti).
At these stages, fertilised eggs can be clearly identified by the development of a blastula.
Nato se lahko za vsako preskusno koncentracijo (tretirano skupino) izračuna odstotek zaviranja hitrosti rasti (Ir) po naslednji enačbi:
Twenty embryos per concentration (one embryo per well) are exposed to the test chemical.
% Ir odstotek zaviranja povprečne specifične hitrosti rasti,
Exposure should be such that ± 20 % of the nominal chemical concentration are maintained throughout the test.
μC aritmetično povprečje za μ v kontroli,
If this is not possible in a static system, a manageable semi-static renewal interval should be applied (e.g. renewal every 24 hrs).
μT aritmetično povprečje za μ v tretirani skupini.
In these cases exposure concentrations need to be verified as a minimum in the highest and lowest test concentrations at the beginning and the end of each exposure interval (see paragraph 36).
Učinki na prirast se določijo na osnovi merjene spremenljivke dolžine glavnega poganjka in treh dodatnih merjenih spremenljivk (tj. po možnosti skupna dolžina poganjka, sveža teža, suha teža ali število spiralasto razvrščenih listov), ki se lahko določijo v vsaki preskusni posodi na začetku in na koncu preskusa.
If an exposure concentration of ± 20 % of the nominal concentrations cannot be maintained, all concentrations need to be measured at the beginning and the end of each exposure interval (see paragraph 36).
V primeru sveže teže ali suhe teže se začetna biomasa določi na osnovi vzorca preskusnih organizmov, ki se vzame iz serije, ki se uporablja za inokulacijo preskusnih posod.
Upon renewal, care should be taken that embryos remain covered by a small amount of old test solutions to avoid drying.
Za vsako preskusno koncentracijo in kontrolo se izračuna aritmetično povprečje prirasta in oceni varianca.
The test design can be adapted to meet the testing requirements of specific substances (e.g,. flow-through (24) or passive dosing systems (25) for easily degradable or highly adsorptive substances (29), or others for volatile substances (36)(37)).
Povprečen odstotek zaviranja prirasta ( % Iy) se lahko za vsako tretirano skupino izračuna po naslednji enačbi:
In any case, care should be taken to minimise any stress to the embryos.
% Iy odstotek zmanjšanja prirasta,
Test chambers should be conditioned at least for 24 hrs with the test solutions prior to test initiation.
bC končna biomasa minus začetna biomasa v kontrolni skupini,
Test conditions are summarised in Appendix 2.
bT končna biomasa minus začetna biomasa v tretirani skupini.
Normally, five concentrations of the test chemical spaced by a constant factor not exceeding 2,2 are required to meet statistical requirements.
Podvojitveni čas
Justification should be provided, if fewer than five concentrations are used.
Za določitev podvojitvenega časa (Td) dolžine glavnega poganjka in izpolnjevanje tega merila za veljavnost (glej odstavek 8) se uporabi naslednja enačba skupaj s podatki iz kontrolnih posod:
The highest concentration tested should preferably result in 100 % lethality, and the lowest concentration tested should preferably give no observable effect, as defined in paragraph 28.
Td = ln 2/μ
A range-finding test before the definitive test allows selection of the appropriate concentration range.
Pri tem je μ povprečna specifična hitrost rasti, ki se določi, kot je opisano v odstavkih od 50 do 52.
The range-finding is typically performed using ten embryos per concentration.
Izdelava krivulj koncentracija–odziv
The following instructions refer to performing the test in 24-well plates.
Izrisati je treba krivulje koncentracija–odziv, ki prikazujejo povprečen odstotek zaviranja spremenljivke odziva (Ir ali Iy, ki se izračuna, kot je prikazano v odstavku 53), in krivuljo logaritemsko transformirane koncentracije preskusne kemikalije.
If different test chambers (e.g. small Petri dishes) are used or more concentrations are tested, instructions have to be adjusted accordingly.
Ocena vrednosti ECx
Details and visual instructions for allocation of concentrations across 24-well plates are available in paragraph 27 and Appendix 4, Figure 1.
Ocene vrednosti ECx morajo temeljiti na povprečni specifični hitrosti rasti (ErCx) in prirastu (EyCx), vsaka od teh vrednosti pa mora temeljiti na dolžini glavnega poganjka in morebitnih dodatnih merjenih spremenljivkah (tj. po možnosti skupna dolžina poganjka, sveža teža, suha teža ali število spiralasto razvrščenih listov).
Dilution water controls are required both as negative control and as internal plate controls.
To je posledica dejstva, da nekatere kemikalije drugače vplivajo na dolžino glavnega poganjka in druge merjene spremenljivke.
If more than 1 dead embryo is observed in the internal plate control, the plate is rejected, thus reducing the number of concentrations used to derive the LC50.
Želeni parametri toksičnosti so zato štiri vrednosti ECx za vsako izračunano raven zaviranja x, in sicer ErCx (dolžina glavnega poganjka), ErCx (tj. po možnosti skupna dolžina poganjka, sveža teža, suha teža ali število spiralasto razvrščenih listov), EyCx (dolžina glavnega poganjka) in EyCx (tj. po možnosti skupna dolžina poganjka, sveža teža, suha teža ali število spiralasto razvrščenih listov).
If an entire plate is rejected the ability to evaluate and discern observed effects may become more difficult, especially if the rejected plate is the solvent control plate or a plate in which treated embryos are also affected.
Opozoriti je treba, da vrednosti ECx, izračunane na podlagi teh dveh spremenljivk odziva, niso primerljive, to razliko pa je treba upoštevati pri uporabi rezultatov preskusa.
In the first case the test must be repeated.
Če se upoštevajo preskusni pogoji iz te preskusne metode, bodo vrednosti ECx, ki temeljijo na povprečni specifični hitrosti rasti (ErCx), v večini primerov višje od rezultatov, ki temeljijo na prirastu (EyCx) zaradi matematične osnove teh dveh pristopov.
In the second one the loss of an entire treatment group(s) due to internal control mortality may limit the ability to evaluate effects and determine LC50 values.
Ta razlika se ne sme razlagati kot razlika v občutljivosti obeh spremenljivk odziva, saj enostavno izvira iz različne matematične narave vrednosti.
A positive control at a fixed concentration of 4 mg/l 3,4-dichloroaniline is performed with each egg batch used for testing.
Statistični postopki
In case a solvent is used, an additional group of 20 embryos is exposed to the solvent on a separate 24-well plate, thus serving as a solvent control.
Cilj je kvantificirati odziv na koncentracijo z regresijsko analizo.
To consider the test acceptable, the solvent should be demonstrated to have no significant effects on time to hatch, survival, nor produce any other adverse effects on the embryos (cf. paragraph 8c).
Po linearni transformaciji podatkov o odzivu se lahko uporabi tehtana linearna regresija, na primer v probit ali logaritemskih ali Weibullovih enotah (7), vendar so nelinearni regresijski postopki primernejše tehnike, ki bolje obravnavajo neizogibne nepravilnosti in odstopanja podatkov od enakomernih porazdelitev.
Start of exposure and duration of test
Če se te nepravilnosti približujejo ničelnemu ali popolnemu zaviranju, se lahko s transformacijo povečajo in ovirajo analizo (7).
The test is initiated as soon as possible after fertilisation of the eggs and terminated after 96 hrs of exposure.
Opozoriti je treba, da so standardne metode probit analize, logaritemske analize ali Weibullove transformacije namenjene uporabi pri kvantnih podatkih (npr. za umrljivost ali preživetje), zato jih je treba prilagoditi za uporabo na podatkih o hitrosti rasti ali prirastu.
The embryos should be immersed in the test solutions before cleavage of the blastodisc commences, or, at latest, by the 16 cell-stage.
Specifični postopki za določanje vrednosti ECx na podlagi kontinuiranih podatkov so na voljo v (8), (9) in (10).
To start exposure with minimum delay, at least twice the number of eggs needed per treatment group are randomly selected and transferred into the respective concentrations and controls (e.g. in 100 ml crystallisation dishes; eggs should be fully covered) not later than 90 minutes post fertilisation.
Za vsako spremenljivko odziva, ki se bo analizirala, se izračuna točkovna ocena vrednosti ECx, pri čemer se uporabi razmerje med odzivom in koncentracijo.
Viable fertilised eggs should be separated from unfertilised eggs and be transferred to 24-well plates pre-conditioned for 24 hrs and refilled with 2 ml/well freshly prepared test solutions within 180 minutes post fertilisation.
Kadar je to mogoče, se za vsako oceno določijo 95-odstotne meje zaupanja.
By means of stereomicroscopy (preferably ≥30-fold magnification), fertilised eggs undergoing cleavage and showing no obvious irregularities during cleavage (e.g. asymmetry, vesicle formation) or injuries of the chorion are selected.
Grafično ali statistično je treba oceniti, kako se podatki o odzivu prilegajo regresijskemu modelu.
For egg collection and separation, see Appendix 3, Fig. 1 and 3 and Appendix 4, Fig.
Regresijsko analizo je treba opraviti na odzivih posameznih ponovljenih vzorcev in ne na povprečjih tretiranih skupin.
2.
Ocene EC50 in meje zaupanja se lahko pridobijo tudi z linearno interpolacijo s samovzorčenjem (10), če razpoložljivi regresijski modeli/metode niso primerni za podatke.
Distribution of eggs over the 24-well plates
Za oceno vrednosti LOEC in s tem tudi vrednosti NOEC je treba primerjati povprečja tretiranih skupin s tehnikami analize variance (ANOVA).
Eggs are distributed to well plates in the following numbers (see also Appendix 4, Fig.
Povprečje za vsako koncentracijo se nato primerja s povprečjem kontrole z ustrezno metodo večkratne primerjave ali testa trenda.
1)
Uporabna sta lahko Dunnettov ali Williamsov test (12) (13) (14) (15) (16).
20 eggs on one plate for each test concentration;
Oceniti je treba, ali velja predpostavka homogenosti varianc v analizi variance.
20 eggs as solvent control on one plate (if necessary);
Ta ocena se lahko izvede grafično ali s formalnim preskusom (15).
20 eggs as positive control on one plate;
Primerna testa sta Levenejev ali Bartlettov test.
4 eggs in dilution water as internal plate control on each of the above plates;
Če predpostavka homogenosti varianc ni izpolnjena, se lahko ponekod to popravi z logaritemsko transformacijo podatkov.
24 eggs in dilution water as negative control on one plate.
Če je heterogenost variance izredno velika in je ni mogoče popraviti s transformacijo, je treba proučiti možnosti analize z metodami, kot je na primer test po Jonckheere-Terpstraju.
Apical observations performed on each tested embryo include: coagulation of embryos, lack of somite formation, non-detachment of the tail, and lack of heartbeat (Table 1).
Dodatna navodila o določanju vrednosti NOEC so na voljo v (10).
These observations are used for the determination of lethality: Any positive outcome in one of these observations means that the zebrafish embryo is dead.
Na podlagi nedavnega znanstvenega razvoja se priporoča opuščanje koncepta vrednosti NOEC in njegova nadomestitev z regresijsko določenimi točkovnimi ocenami ECx.
Additionally, hatching is recorded in treatment and control groups on a daily basis starting from 48 hrs.
Za ta preskus Myriophyllum ni bila določena ustrezna vrednost x.
Observations are recorded every 24 hrs, until the end of the test.
Vendar se zdi, da je ustrezno območje od 10 % do 20 % (odvisno od izbrane spremenljivke odziva), po možnosti pa je treba poročati tako o vrednostih EC10 in EC20 kot o njunih mejah zaupanja.
Apical observations of acute toxicity in zebrafish embryos 24-96 hrs post fertilisation
Poročanje
Exposure times
Snov z več sestavinami, UVCB ali zmes:
24 hrs
Preskusne vrste
48 hrs
Znanstveno ime in vir.
72 hrs
Uporabljeni preskusni postopek (statični ali polstatični).
96 hrs
Datum začetka preskusa in njegovo trajanje.
Coagulated embryos
Preskusno gojišče.
Lack of somite formation
Opis načrta poskusa: preskusne posode in pokrovi, količine raztopin in dolžina glavnega poganjka za vsako poskusno posodo na začetku preskusa.
Non-detachment of the tail
Preskusne koncentracije (nominalne in izmerjene, kot je ustrezno) in število ponovljenih vzorcev za posamezno koncentracijo.
Lack of heartbeat
Metode priprave osnovnih in testnih raztopin, vključno z uporabo kakršnih kolih topil ali disperzijskih sredstev.
Coagulation of the embryo: Coagulated embryos are milky white and appear dark under the microscope (see Appendix 5, Fig.
Temperatura med preskusom.
1).
Svetlobni vir, obsevanost in homogenost.
The number of coagulated embryos is determined after 24, 48, 72 and 96 hrs.
Vrednosti pH preskusnega in kontrolnega gojišča.
Lack of somite formation: At 26 ± 1 °C, about 20 somites have formed after 24 hrs (see Appendix 5, Figure 2) in a normally developing zebrafish embryo.
Metoda analize preskusne kemikalije z ustreznimi podatki o oceni kakovosti (validacijske študije, standardni odkloni ali meje zaupanja analiz).
A normally developed embryo shows spontaneous movements (side-to-side contractions).
Metode za določanje dolžine glavnega poganjka in drugih merjenih spremenljivk, npr. skupne dolžine stranskih vej, skupne dolžine poganjka, skupne dolžine korenin, sveže teže, suhe teže in števila spiralasto razvrščenih listov.
Non-formation of somites after 24 hrs might be due to a general retardation of development.
Neobdelani podatki: dolžina glavnega poganjka in druge merjene spremenljivke v vsaki preskusni in kontrolni posodi pri vsakem opazovanju in analizi.
At latest after 48 hrs, the formation of somites should be developed.
Povprečja in standardni odkloni za vsako merjeno spremenljivko.
If not, the embryos are considered dead.
Krivulje rasti za vsako merjeno spremenljivko.
Non-detachment of the tail: In a normally developing zebrafish embryo, detachment of the tail (seeAppendix 5, Figure 3) from the yolk is observed following posterior elongation of the embryonic body.
Izračunane spremenljivke odziva za vsak tretirani ponovljeni vzorec z aritmetičnimi povprečji in koeficientom variacije za ponovljene vzorce.
Absence of tail detachment is recorded after 24, 48, 72 and 96 hrs.
Grafična predstavitev razmerja med koncentracijo in učinkom.
Lack of heartbeat: In a normally developing zebrafish embryo at 26 ± 1 °C, the heartbeat is visible after 48 hrs (see Appendix 5, Figure 4).
Če je bila uporabljena analiza variance, velikost opaženega učinka (npr. najmanjša značilna razlika).
Particular care should be taken when recording this endpoint, since irregular (erratic) heartbeat should not be recorded as lethal.
Vsaka stimulacija rasti, opažena pri katerem koli tretiranju.
Moreover, visible heartbeat without circulation in aorta abdominalis is considered non-lethal.
Vsi vidni znaki fitotoksičnosti in opažanja testnih raztopin.
To record this endpoint, embryos showing no heartbeat should be observed under a minimum magnification of 80x for at least one minute.
Razprava o rezultatih, vključno z vsemi vplivi na rezultat preskusa, ki izhajajo iz odstopanj od te preskusne metode.
Absence of heartbeat is recorded after 48, 72 and 96 hrs.
ASTM Designation E 1913-04.
Hatching rates of all treatment and control groups should be recorded from 48 hrs onwards and reported.
Standard Guide for Conducting Static, Axenic, 14-Day Phytotoxicity Tests in Test Tubes with the Submersed Aquatic Macrophyte, Myriophyllum sibiricum Komarov.
Although hatching is not an endpoint used for the calculation of the LC50, hatching ensures exposure of the embryo without a potential barrier function of the chorion, and as such may help data interpretation.
(1) Maletzki, D. et al.
Detailed descriptions of the normal (35) and examples of abnormal development of zebrafish embryos are illustrated in Appendixes 3 and 5.
Myriophyllum spicatum als ökotoxikologischer Testorganismus: Methodenentwicklung eines sedimentfreien Testsystems und erste Ergebnisse mit 3,5-Dichlorphenol, Umweltwiss Schadst Forsch, št. 22, str. 702–710.
Analytical measurements
Poglavje C.26 te priloge: Preskus zaviranja rasti Lemna sp.
At the beginning and at the end of the test, pH, total hardness and conductivity in the control(s) and in the highest test chemical concentration are measured.
OECD (2014).
In semi-static renewal systems the pH should be measured prior to and after water renewal.
„Myriophyllum spicatum Toxicity Test: Results of an inter-laboratory ring test using a sediment-free test system“, OECD Environment, Health and Safety Publications (EHS), Series Testing and Assessment, št. 205, OECD Publishing, Pariz.
The dissolved oxygen concentration is measured at the end of the test in the negative controls and highest test concentration with viable embryos, where it should be in compliance with the test validity criteria (see paragraph 8f).
(4) OECD (2000). „Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures“, OECD Environment, Health and Safety Publications (EHS), Series on Testing and Assessment, št. 23, OECD Publishing, Pariz.
If there is concern that the temperature varies across the 24-well plates, temperature is measured in three randomly selected vessels.
OECD (2000). „Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures“, OECD Environment, Health and Safety Publications (EHS), Series on Testing and Assessment, št. 23, OECD Publishing, Pariz.
Temperature should be recorded preferably continuously during the test or, as a minimum, daily.
Christensen, E. R., N. Nyholm (1984).
In a static system, the concentration of the test chemical should be measured, as a minimum, in the highest and lowest test concentrations, but preferably in all treatments, at the beginning and end of the test.
Ecotoxicological Assays with Algae: Weibull Dose-Response Curves, Environmental Science & Technology, zv.
In semi-static (renewal) tests where the concentration of the test chemical is expected to remain within ± 20 % of the nominal values, it is recommended that, as a minimum, the highest and lowest test concentrations be analysed when freshly prepared and immediately prior to renewal.
18/9, str. 713–718.
For tests where the concentration of the test chemical is not expected to remain within ± 20 % of nominal, all test concentrations must be analysed when freshly prepared and immediately prior to renewal.
(7) Nyholm, N. et al.
In case of insufficient volume for analysis, merging of test solutions, or use of surrogate chambers being of the same material and having the same volume to surface area ratios as 24-well plates, may be useful.
Statistical treatment of data from microbial toxicity tests, Environmental Toxicology and Chemistry, zv.
It is strongly recommended that results be based on measured concentrations.
Bruce, R. D., D. J. Versteeg (1992).
When the concentrations do not remain within 80-120 % of the nominal concentration, the effect concentrations should be expressed relative to the geometric mean of the measured concentrations; see Chapter 5 in the OECD Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures for more details (28).
A statistical procedure for modelling continuous toxicity data, Environmental Toxicology and Chemistry, zv.
LIMIT TEST
OECD (2006). „Current Approaches in the Statistical Analysis of Ecotoxicity Data: A Guidance to Application“, OECD Environment, Health and Safety Publications (EHS), Series on Testing and Assessment, št. 54, OECD Publishing, Pariz.
Using the procedures described in this test method, a limit test may be performed at 100 mg/l of test chemical or at its limit of solubility in the test medium (whichever is the lower) in order to demonstrate that the LC50 is greater than this concentration.
Norberg-King, T. J. (1988).
The limit test should be performed using 20 embryos in the treatment, the positive control and –if necessary- in the solvent control and 24 embryos in the negative control.
Dunnett, C. W.
If the percentage of lethality at the concentration tested exceeds the mortality in the negative control (or solvent control) by 10 %, a full study should be conducted.
A multiple comparisons procedure for comparing several treatments with a control, Journal of the American Statistical Association, zv.
Any observed effects should be recorded.
50/272, str. 1096–1121.
If mortality exceeds 10 % in the negative control (or solvent control), the test becomes invalid and should be repeated.
New tables for multiple comparisons with a control, Biometrics, zv.
In this test, the individual wells are considered independent replicates for statistical analysis.
20/3, str. 482-491.
The percentages of embryos for which at least one of the apical observations is positive at 48 and/or 96 hrs are plotted against test concentrations.
Williams, D. A. (1972).
For calculation of the slopes of the curve, LC50 values and the confidence limits (95 %), appropriate statistical methods should be applied (38) and the OECD Guidance Document on Current Approaches in the Statistical Analysis of Ecotoxicity Data should be consulted (39).
The comparison of several dose levels with a zero dose control, Biometrics, zv.
chemical identification, such as IUPAC or CAS name, CAS number, SMILES or InChI code, structural formula, purity, chemical identity of impurities as appropriate and practically feasible, etc. (including the organic carbon content, if appropriate).
27/1, str. 103–117.
Test organisms:
28/2, str. 519–531.
test procedure used (e.g., semi-static renewal);
29/2, str. 93–96.
photoperiod;
Biomasa je sveža in/ali suha teža žive snovi, prisotne v populaciji.
test design (e.g., number of test chambers, types of controls);
Pri tem preskusu je biomasa vsota glavnega poganjka, vseh stranskih vej in vseh korenin.
water quality characteristics in fish maintenance (e.g. pH, hardness, temperature, conductivity, dissolved oxygen);
Kloroza je sprememba barve preskusnega organizma, zlasti spiralasto razvrščenih listov, z zelene na rumenkasto.