natgillin/translations-raw huggingface dataset:
Glass or polystyrene test chambers should be used (e.g. 24-well plates with a 2,5-5 ml filling capacity per well).
podaci za kemijsku identifikaciju tvari, kao što su IUPAC ili CAS naziv, CAS broj, oznaka SMILES ili InChI, strukturna formula, čistoća, kemijski identitet nečistoća prema potrebi i ako je praktički izvedivo itd. (uključujući udio organskog ugljika, prema potrebi),
In case adsorption to polystyrene is suspected (e.g., for non-polar, planar substances with high KOW), inert materials (glass) should be used to reduce losses due to adsorption (26).
Ispitni organizmi:
Test chambers should be randomly positioned in the incubator.
ispitni postupak (npr. polustatičko ispitivanje s obnavljanjem),
Water and test conditions
fotoperiod,
Dilution of the maintenance water is recommended to achieve hardness levels typical of a wide variety of surface waters.
plan ispitivanja (npr. broj ispitnih komora, vrste kontrole),
Dilution water should be prepared from reconstituted water (27).
obilježja kvalitete vode za držanje riba (npr. pH, tvrdoća, temperatura, vodljivost, otopljeni kisik),
The resulting degree of hardness should be equivalent to 100-300 mg/l CaCO3 in order to prevent excessive precipitation of calcium carbonate.
koncentracija otopljenog kisika, pH, ukupna tvrdoća, temperatura i vodljivost ispitnih otopina na početku i nakon 96 sati,
Other well-characterised surface or well water may be used.
način pripreme matičnih otopina i ispitnih otopina te učestalost obnavljanja,
The reconstituted water may be adapted to maintenance water of low hardness by dilution with deionised water up to a ratio of 1:5 to a minimum hardness of 30-35 mg/l CaCO3.
obrazloženje za upotrebu otapala i obrazloženje za odabir otapala (ako nije voda),
The water is aerated to oxygen saturation prior to addition of the test chemical.
nazivne ispitne koncentracije i rezultati svih analiza za određivanje koncentracije ispitivane kemikalije u ispitnim posudama; trebalo bi navesti i učinkovitost izdvajanja ispitivane tvari pri primjeni metode te granicu kvantifikacije (LoQ),
Temperature should be kept at 26 ± 1 °C, in the wells, throughout the test.
dokaz da kontrole ispunjavaju opće kriterije valjanosti za preživljavanje,
The pH should be in a range between pH 6,5 and 8,5, and not vary within this range by more than 1,5 units during the course of the test.
stopa oplodnje jajašaca,
If the pH is not expected to remain in this range, then pH adjustment should be done prior to initiating the test.
stopa valjenja u tretiranim i kontrolnim skupinama.
The pH adjustment should be made in such a way that the stock solution concentration is not changed to any significant extent and that no chemical reaction or precipitation of the test chemical is caused.
najviša koncentracija koja ne izaziva smrtnost u razdoblju ispitivanja,
Use of hydrogen chloride (HCl) and sodium hydroxide (NaOH) to correct pH in the solutions containing the test chemical is recommended.
najniža koncentracija koja izaziva 100 %-tnu smrtnost u razdoblju ispitivanja,
Test solutions of the selected concentrations can be prepared, e.g. by dilution of a stock solution.
ukupna smrtnost u svakoj koncentraciji u svakom preporučenom vremenu promatranja,
The stock solutions should preferably be prepared by simply mixing or agitating the test chemical in the dilution water by mechanical means (e.g. stirring and/or ultra-sonification).
vrijednosti LC50 nakon 96 sati (i eventualno nakon 48 sati) za smrtnost s granicama pouzdanosti od 95 %, ako je to moguće,
If the test chemical is difficult to dissolve in water, procedures described in the OECD Guidance Document No. 23 for handling difficult substances and mixtures should be followed (28).
graf krivulje za odnos koncentracija i smrtnosti na kraju ispitivanja,
Where a solvent is used to assist in stock solution preparation, its final concentration should not exceed 100 μl/l and should be the same in all test vessels.
podaci o ishodima svakog od četiri apikalna zapažanja,
When a solvent is used, an additional solvent control is required.
pojava i opis morfoloških i fizioloških anomalija, ako ih ima (vidjeti primjere navedene u Dodatku 5. slici 2.),
Maintenance of brood fish
svi događaji tijekom ispitivanja koji su možda utjecali na rezultate,
A breeding stock of unexposed, wild-type zebrafish with well-documented fertilisation rate of eggs is used for egg production.
statistička analiza i obrada podataka (probit analiza, logistički regresijski model i geometrijska sredina za LC50),
Fish should be free of macroscopically discernible symptoms of infection and disease and should not have undergone any pharmaceutical (acute or prophylactic) treatment for 2 months before spawning.
nagib i granice pouzdanosti regresije (transformirane) krivulje koncentracija-odgovor.
Breeding fish are maintained in aquaria with a recommended loading capacity of 1 l water per fish and a fixed 12 - 16 hour photoperiod (29)(30)(31)(32)(33).
Sva odstupanja od ispitne metode i odgovarajuća objašnjenja.
Optimal filtering rates should be adjusted; excess filtering rates causing heavy perturbation of the water should be avoided.
Diskusija i tumačenje rezultata.
For feeding conditions, see Appendix 2.
OECD (2011) Validation Report (Phase 1) for the Zebrafish Embryo Toxicity Test: Part I and Part II.
Surplus feeding should be avoided, and water quality and cleanness of the aquaria should be monitored regularly and be reset to the initial state, if necessary.
Series on Testing and Assessment No.
Proficiency Testing
157, OECD, Paris.
As a reference chemical, 3,4-dichloroaniline (used in the validation studies (1)(2)), should be tested in a full concentration-response range to check the sensitivity of the fish strain used, preferably twice a year.
OECD (2012) Validation Report (Phase 2) for the Zebrafish Embryo Toxicity Test: Part I and Part II (Annexes).
For any laboratory initially establishing this assay, the reference chemical should be used.
Series on Testing and Assessment No.
A laboratory can use this chemical to demonstrate their technical competence in performing the assay prior to submitting data for regulatory purposes.
179, OECD, Paris.
Egg production
Environmental Toxicology and Chemistry 32: 1768-1783.
Zebrafish eggs may be produced via spawning groups (in individual spawning tanks) or via mass spawning (in the maintenance tanks).
Pharmacol.: 149 (2), 196-209.
Since spawning groups of zebrafish may occasionally fail to spawn, the parallel use of at least three spawning tanks is recommended.
Poglavlje C.29. ovog Priloga: Laka biorazgradivost – CO2 u začepljenim posudama.
To avoid genetic bias, eggs are collected from a minimum of three breeding groups, mixed and randomly selected.
Poglavlje C.48. ovog Priloga: Kratkoročni reprodukcijski test na ribama.
To prevent predation of eggs by adult zebrafish, the spawn traps are covered with inert wire mesh of appropriate mesh size (approx.
35, 4097-4102.
2 ± 0,5 mm).
Dostupno na: [http://www.iso.org].
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).
23, OECD, Paris.
Weathered plastic materials which do not leach (e.g., phthalates) should be used.
Europska komisija (2007.): Preporuka Komisije 2007/526/EZ od 18. lipnja 2007. o smjernicama za smještaj životinja kojima se koristi za pokusne i druge znanstvene svrhe te skrb o njima (priopćena pod brojem dokumenta C(2007) 2525) [http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2007:197:0001:0089:EN:PDF].
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.
Europska unija (2010.): Direktiva 2010/63/EU Europskog parlamenta i Vijeća od 22. rujna 2010. o zaštiti životinja koje se koriste u znanstvene svrhe.
Rinsing eggs with reconstituted water after collection from spawning traps is recommended.
Službeni list Europske unije, L 276: 33. – 79.; 20.10.2010.
Egg differentiation
http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2010:276:0033:0079:EN:PDF.
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).
Poglavlje C.2. ovog Priloga: Test akutne imobilizacije Daphnia sp.
At these stages, fertilised eggs can be clearly identified by the development of a blastula.
Series on Testing and Assessment No.
Twenty embryos per concentration (one embryo per well) are exposed to the test chemical.
OECD, Paris.
Exposure should be such that ± 20 % of the nominal chemical concentration are maintained throughout the test.
Detaljni pregled ‚Fish embryo toxicity assays’.
If this is not possible in a static system, a manageable semi-static renewal interval should be applied (e.g. renewal every 24 hrs).
Apikalna krajnja točka
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).
uzrokuje učinke na razini populacije.
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).
stanična formacija oko animalnog pola koja obuhvaća određeni dio žumanjka.
Upon renewal, care should be taken that embryos remain covered by a small amount of old test solutions to avoid drying.
tvar ili smjesa.
In any case, care should be taken to minimise any stress to the embryos.
veliko umnožavanje pretežno epidermalnih stanica u fazi gastrulacije embrija i njihovo premještanje s dorzalne na ventralnu stranu, čime se entodermalni slojevi stanica uvlače u procesu sličnom invaginaciji i žumanjak se ugrađuje u embrij.
Test chambers should be conditioned at least for 24 hrs with the test solutions prior to test initiation.
Protočno ispitivanje
Test conditions are summarised in Appendix 2.
test sa stalnim protokom ispitne otopine kroz ispitni sustav za vrijeme izlaganja.
Normally, five concentrations of the test chemical spaced by a constant factor not exceeding 2,2 are required to meet statistical requirements.
Kontrola unutar ploče
Justification should be provided, if fewer than five concentrations are used.
unutarnja kontrola koja se sastoji od 4 bunarića napunjena vodom za razrjeđivanje po ploči s 24 bunarića radi utvrđivanja moguće kontaminacije ploča koju prouzroči proizvođač ili istraživač tijekom postupka i svih učinaka na ploču koji mogu utjecati na rezultat ispitivanja (npr. promjena temperature).
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.
Međunarodna unija za čistu i primijenjenu kemiju
A range-finding test before the definitive test allows selection of the appropriate concentration range.
Voda za držanje
The range-finding is typically performed using ten embryos per concentration.
voda u kojoj se drže odrasle ribe.
The following instructions refer to performing the test in 24-well plates.
Medijan smrtonosne koncentracije (LC50)
If different test chambers (e.g. small Petri dishes) are used or more concentrations are tested, instructions have to be adjusted accordingly.
koncentracija ispitivane kemikalije koja je procijenjena kao smrtonosna za 50 % ispitnih organizama za vrijeme ispitivanja.
Details and visual instructions for allocation of concentrations across 24-well plates are available in paragraph 27 and Appendix 4, Figure 1.
Polustatičko ispitivanje s obnavljanjem
Dilution water controls are required both as negative control and as internal plate controls.
ispitivanje s redovitim obnavljanjem ispitne otopine nakon određenih razdoblja (npr. svaka 24 sata).
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.
Somit
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.
u embriju kralježnjaka koji se razvija somiti su nakupine mezoderma raspoređene na lateralnom dijelu neuralne cijevi iz kojih se s vremenom razvijaju dermis (dermatom), skeletni mišići (miotom) i kralješci (sklerotom).
In the first case the test must be repeated.
Statičko ispitivanje
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.
ispitivanje u kojem ispitne otopine ostaju nepromijenjene tijekom cijelog ispitivanja.
A positive control at a fixed concentration of 4 mg/l 3,4-dichloroaniline is performed with each egg batch used for testing.
bilo koja tvar ili smjesa koja se ispituje ovom ispitnom metodom.
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.
tvari nepoznatog ili promjenjivog sastava, složeni reakcijski proizvodi i biološki materijali.
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).
DRŽANJE, UZGOJ I TIPIČNI UVJETI ZA ISPITIVANJA AKUTNE TOKSIČNOSTI NA EMBRIJIMA ZEBRICA
Start of exposure and duration of test
Zebrica (Danio rerio)
The test is initiated as soon as possible after fertilisation of the eggs and terminated after 96 hrs of exposure.
Podrijetlo vrste
The embryos should be immersed in the test solutions before cleavage of the blastodisc commences, or, at latest, by the 16 cell-stage.
Indija, Burma, Melaka, Sumatra
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.
Spolni dimorfizam
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.
Ženke: izbočen trbuh kada nose jajašca
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.
Mužjaci: tanji, narančasta nijansa između plavih vodoravnih pruga (posebno očita na analnoj peraji)
For egg collection and separation, see Appendix 3, Fig. 1 and 3 and Appendix 4, Fig.
Suha hrana u listićima (najviše 3 % težine ribe po danu) 3 – 5 puta dnevno; usto ličinke (nauplii) račića Artemia i/ili male vodenbuhe odgovarajuće veličine dobivene iz nekontaminiranih izvora.
2.
Živom se hranom bogati okoliš i stoga bi kad god je to moguće trebalo davati živu hranu.
Distribution of eggs over the 24-well plates
Kako bi se osigurala optimalna kvaliteta vode, trebalo bi ukloniti višak hrane i izmet otprilike jedan sat nakon hranjenja.
Eggs are distributed to well plates in the following numbers (see also Appendix 4, Fig.
Približna težina odraslih riba
1)
Ženke: 0,65 ± 0,13 g
20 eggs on one plate for each test concentration;
Mužjaci: 0,5 ± 0,1 g
20 eggs as solvent control on one plate (if necessary);
Držanje uzgojnih riba
20 eggs as positive control on one plate;
Osvjetljenje
4 eggs in dilution water as internal plate control on each of the above plates;
Fluorescentne žarulje (širokog spektra); 10 – 20 μE/m2/s, 540 – 1080 lux, ili 50 – 100 ft-c (razine u laboratorijskim uvjetima); fotoperiod 12 – 16 sati
24 eggs in dilution water as negative control on one plate.
26 ± 1 °C
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).
Kvaliteta vode
These observations are used for the determination of lethality: Any positive outcome in one of these observations means that the zebrafish embryo is dead.
Zasićenost O2 ≥ 80 %, tvrdoća: npr. ~30 – 300 mg/l CaCO3, NO3–: ≤ 48 mg/l, NH4+ i NO2–: < 0,001 mg/l, rezidualni klor < 10 μg/l, ukupni organski klor < 25 ng/l, pH = 6,5 – 8,5
Additionally, hatching is recorded in treatment and control groups on a daily basis starting from 48 hrs.
Dodatni kriteriji za kvalitetu vode
Observations are recorded every 24 hrs, until the end of the test.
Čestice < 20 mg/l, ukupni organski ugljik < 2 mg/l, ukupni organofosforni pesticidi < 50 ng/l, ukupni organoklorni pesticidi plus poliklorirani bifenili < 50 ng/l
Apical observations of acute toxicity in zebrafish embryos 24-96 hrs post fertilisation
Veličina akvarija za držanje
Exposure times
Npr.
24 hrs
180 l, 1 riba/l
48 hrs
Pročišćavanje vode
72 hrs
Stalno (filtriranje ugljenom); druge mogućnosti uključuju kombinacije s polustatičkim sustavom s obnavljanjem ili protočnim sustavom s neprekidnim obnavljanjem vode
96 hrs
Preporučeni omjer mužjaka i ženki za razmnožavanje
Coagulated embryos
2: 1 (ili masovno mriješćenje)
Lack of somite formation
Akvariji za mriješćenje
Non-detachment of the tail
Npr. akvariji od 4 l s dnom od čelične mreže i umjetnom biljkom kao podražajem za mriješćenje; vanjski grijači, ili masovno mriješćenje u akvarijima za držanje
Lack of heartbeat
Struktura i izgled jajašaca
Coagulation of the embryo: Coagulated embryos are milky white and appear dark under the microscope (see Appendix 5, Fig.
Stabilan korion (tj. vrlo proziran, neljepljiv, promjera ~ 0,8 – 1,5 mm)
1).
Stopa mriješćenja
The number of coagulated embryos is determined after 24, 48, 72 and 96 hrs.
Jedna zrela ženka mrijesti najmanje 50 – 80 jajašaca dnevno.
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.
Ovisno o soju, stopa mriješćenja može biti znatno viša.
A normally developed embryo shows spontaneous movements (side-to-side contractions).
Stopa oplodnje trebala bi biti ≥ 70 %.
If not, the embryos are considered dead.
Odabrane faze ranog razvoja zebrice (Danio rerio): 0,2 – 1,75 sati nakon oplodnje (iz: Kimmel et al.
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.
1995 (35.)).
Absence of tail detachment is recorded after 24, 48, 72 and 96 hrs.
S pomoću vremenskog slijeda normalnog razvoja mogu se dijagnosticirati i oplodnja i iskoristivost jajašaca (vidjeti stavak 26.: Odabir oplođenih jajašaca).
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).
Slika 2.
Particular care should be taken when recording this endpoint, since irregular (erratic) heartbeat should not be recorded as lethal.
Odabrane faze kasnog razvoja zebrice (Danio rerio) (embrij bez koriona radi bolje vidljivosti): 22 sata – 48 sati nakon oplodnje (iz: Kimmel et al.
Moreover, visible heartbeat without circulation in aorta abdominalis is considered non-lethal.
1995 (35.)).
To record this endpoint, embryos showing no heartbeat should be observed under a minimum magnification of 80x for at least one minute.
Slika 3.
Absence of heartbeat is recorded after 48, 72 and 96 hrs.
Normalan razvoj embrija zebrice (Danio rerio): (1) 0,75 sati, faza s 2 stanice; (2) 1 sat, faza s 4 stanice; (3) 1,2 sata, faza s 8 stanica; (4) 1,5 sati, faza s 16 stanica; (5) 4,7 sati, početak epibolije; (6) 5,3 sata, otprilike 50 % epibolije (iz: Braunbeck & Lammer 2006 (40.)).
Hatching rates of all treatment and control groups should be recorded from 48 hrs onwards and reported.
Nacrt ploča s 24 bunarića
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 – 5 pet ispitnih koncentracija po kemikaliji;
Detailed descriptions of the normal (35) and examples of abnormal development of zebrafish embryos are illustrated in Appendixes 3 and 5.
nC negativna kontrola (voda za razrjeđivanje);
Analytical measurements
iC kontrola unutar ploče (voda za razrjeđivanje);
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.
pC pozitivna kontrola (3,4-dikloroanilin 4 mg/l);
In semi-static renewal systems the pH should be measured prior to and after water renewal.
sC kontrola s otapalom
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).
PRIKAZ SMRTNIH KRAJNJIH TOČAKA ZA ISPITIVANJE AKUTNE TOKSIČNOSTI NA EMBRIJIMA ZEBRICA
If there is concern that the temperature varies across the 24-well plates, temperature is measured in three randomly selected vessels.
Sljedeće apikalne krajnje točke znak su akutne toksičnosti te, posljedično, smrti embrija: koagulacija embrija, neodvajanje repa, izostanak formiranja somita i izostanak otkucaja srca.
Temperature should be recorded preferably continuously during the test or, as a minimum, daily.
Sljedeće mikrofotografije odabrane su za ilustraciju tih krajnjih točaka.
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.
Koagulacija embrija
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.
Pri promatranju uz jako osvjetljenje polja na koaguliranim embrijima zebrica vidljiv je niz neprozirnih nakupina.
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.
Slika 2.
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.
Izostanak formiranja somita
It is strongly recommended that results be based on measured concentrations.
Unatoč kašnjenju u razvoju za otprilike 10 sati, u embrija zebrice starog 24 sata na slici a) vide se dobro razvijeni somiti (→), dok u embrija na slici b) nema nikakvog znaka formiranja somita (→).
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).
Unatoč velikom edemu žumanjčane vrećice (*), u embrija zebrice starog 48 sati na slici c) dobro se vide formirani somiti (→), dok u embrija zebrice starog 96 sati na slici d) nema nikakvog znaka formiranja somita (→).
LIMIT TEST
U embrija na slici d) zamjetni su i zakrivljenost kralježnice (skolioza) i perikardijalni edem (*).
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.
(a: →; embrij zebrice star 96 sati).
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.
Zamjetno je i da nema osnove oka (*).
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.
Slika 4.