natgillin/translations-raw huggingface dataset:
It is especially recommended to make such a check to ensure that the performance of the test is not compromised if the organic carbon content of the artificial soil is reduced, e.g. by lowering the peat content to 4-5 % and increasing the sand content accordingly.
Priporočljiva je metoda, ki temelji na barvanju z barvilom bengal rdeče (glej Dodatek 6), čeprav so se kot primerne izkazale tudi druge tehnike ekstrakcije z vlago (ne pa tudi s toploto) in flotacije (glej Dodatek 6) (4)(10)(11)(20).
By such a reduction in organic carbon content, the possibilities of adsorption of test chemical to the soil (organic carbon) may be decreased and the availability of the test chemical to the worms may increase.
Barvanje z barvilom bengal rdeče je priporočljivo, ker ekstrakcijo iz substrata tal z vlago lahko ovira motnost zaradi delcev suspendirane gline.
It has been demonstrated that Enchytraeus albidus can comply with the validity criteria on reproduction when tested in field soils with lower organic carbon content than mentioned above (e.g. 2,7 %) (15), and there is experience — though limited — that this can also be achieved in artificial soil with 5 % peat.
Če pri največji koncentraciji v preskusu za določanje območja delovanja (tj. 1000 mg/kg) ni opaženih učinkov, je mogoče preskus razmnoževanja s koncentracijo 1000 mg/kg izvesti kot mejni preskus, da se dokaže, da je vrednost NOEC za razmnoževanje večja od te vrednosti.
When using natural soil in additional (e.g. higher tier) testing, the suitability of the soil and achieving the test validity criteria should also be demonstrated.
Povzetek in časovni razpored preskusa
The dry constituents of the soil are mixed thoroughly (e.g. in a large-scale laboratory mixer).
Koraki preskusa so naslednji:
This should be done at least one week before starting the test.
Čas
The mixed soil should be stored for two days in order to equilibrate/stabilise the acidity.
dan – 7 ali prej
For the determination of pH a mixture of soil and 1 M potassium chloride (KCl) or 0,01 M calcium chloride (CaCl2) solution in a 1:5 ratio is used (see (16) and Appendix 3).
priprava umetnih tal (mešanje suhih sestavin);
If the soil is more acidic than the required range (see paragraph 11), it can be adjusted by addition of an appropriate amount of CaCO3.
dan – 5
If the soil is too alkaline it can be adjusted by the addition of more of the mixture, referred to in paragraph 11, but excluding the CaCO3.
preverjanje vrednosti pH umetnih tal
The maximum water holding capacity (WHC) of the artificial soil is determined in accordance with procedures described in Appendix 2.
meritev največje zmogljivosti tal za zadrževanje vode;
One or two days before starting the test, the dry artificial soil is pre-moistened by adding enough de-ionised water to obtain approximately half of the final water content, that being 40 to 60 % of the maximum water holding capacity.
od dne – 5 do dne – 3
At the start of the test, the pre-moistened soil is divided into portions corresponding with the number of test concentrations (and reference chemical where appropriate) and controls used for the test.
razvrščanje črvov za aklimatizacijo
The moisture content is adjusted to 40-60 % of the maximum WHC by the addition of the test chemical solution and/or by adding distilled or de-ionised water (see paragraphs 19-21).
od dne – 3 do dne 0
The moisture content is determined at the beginning and at the end of the test (by drying to constant weight at 105 °C) and should be within the optimal range for the survival of the worms.
aklimatizacija črvov vsaj 24 ur
A rough check of the soil moisture content can be obtained by gently squeezing the soil in the hand, if the moisture content is correct small drops of water should appear between the fingers.
dan – 1
Selection and preparation of test animals
predhodna navlažitev umetnih tal in razdelitev v šarže
The recommended test species is Enchytraeus albidus Henle 1837 (white potworm), a member of the family Enchytraeidae (order Oligochaeta, phylum Annelida).
Dan 0
E. albidus is one of the largest species of enchytraeids, with specimens of up to 35 mm in length being recorded (17)(18).
priprava založnih raztopin
E. albidus has a world-wide distribution and is found in marine, freshwater and terrestrial habitats, mainly in decaying organic matter (seaweed, compost) and rarely in meadows (9).
nanos preskusne kemikalije
E. albidus is commercially available, as a fish food.
primešanje hrane
It should be checked whether the culture is contaminated by other, usually smaller, species (1) (19).
vnašanje črvov
If contamination occurs, all worms should be washed with water in a petri dish.
merjenje vrednosti pH in vsebnosti vlage v tleh
Large adult specimens of E. albidus should then be selected (using a stereomicroscope) to start a new culture and all other worms are discarded.
dan
E. albidus can be bred easily in a wide range of organic materials (see Appendix 4).
preverjanje vsebnosti vlage v tleh
The life-cycle of E. albidus is short since maturity is reached between 33 days (at 18 °C) and 74 days (at 12 °C) (1).
hranjenje
Only cultures that have been kept without problems in the laboratory for at least 5 weeks (one generation) will be used for the test.
Dan 14
Other species of the Enchytraeus genus are also suitable, e.g. E. buchholzi Vejdovsky 1879 or E. crypticus Westheide & Graefe 1992 (see Appendix 5).
določitev smrtnosti odraslih osebkov
If other species of Enchytraeus are used, they must be clearly identified and the rationale for the selection of the species should be reported.
ocena števila mladičev
The animals used in the tests are adult worms.
dan
They should have eggs (white spots) in the clitellum region, and they should be approximately the same size (about 1 cm long).
opazovanje vedenja odraslih osebkov
Synchronisation of the breeding culture is not necessary.
odstranjevanje odraslih osebkov
If the enchytraeids are not bred in the same soil type and under the conditions (including feeding) used for the final test they must be acclimatised for at least 24 hours and up to three days.
dan 28
A larger number of adults than that needed for performing the test should initially be acclimatised to allow scope for rejection of damaged or otherwise unsuitable specimens.
brez hranjenja
At the end of the acclimatisation period, only worms containing eggs and exhibiting no behavioural abnormalities (e.g. trying to escape from the soil) are selected for the test.
dan 35
The worms are carefully removed using jeweller's tweezers, hooks or loops and placed in a petri dish containing a small amount of fresh water.
dan 42
Reconstituted fresh water as proposed in Chapter C.20 of this Annex (Daphnia magna Reproduction Test) is preferred for this purpose since de-ionised, de-mineralised or tap water could be harmful to the worms.
štetje mladih črvov
The worms are inspected under a stereomicroscope and any that do not contain eggs are discarded.
Obdelava rezultatov
Care is taken to remove and discard any mites or springtails that might have infected the cultures.
Čeprav je v Dodatku 7 podan pregled, v tej preskusni metodi ni podana nobena dokončna statistična smernica za analizo rezultatov preskusa.
Healthy worms not used for the test are returned to the stock culture.
V preskusu za določanje območja delovanja je glavna končna točka smrtnost.
Preparation of test concentrations
Vendar je treba zabeležiti tudi spremembe vedenja odraslih osebkov (npr. nezmožnost kopanja v tla, negibno ležanje ob stekleni steni preskusne posode) in morfologije (npr. odprte rane) ter morebitno prisotnost mladičev.
Test chemical soluble in water
Za določanje vrednosti LC50 se običajno uporabi analiza probit (21) ali regresija logit.
A solution of the test chemical is prepared in deionised water in a quantity sufficient for all replicates of one test concentration.
Vendar pa je mogoče v primerih, pri katerih ta analitska metoda ni primerna (npr. če so na voljo manj kot tri koncentracije z delnimi usmrtitvami), uporabiti alternativne metode.
It is recommended to use an appropriate quantity of water to reach the required moisture content, i.e. 40 to 60 % of the maximum WHC (see paragraph 13).
Te metode lahko vključujejo drseča povprečja (22), prilagojeno metodo Spearman-Karber (23) ali enostavno interpolacijo (npr. geometrijsko sredino vrednosti LC0 in LC100, izračunano s kvadratnim korenom zmnožka vrednosti LC0 in LC100).
Each solution of test chemical is mixed thoroughly with one batch of pre-moistened soil before being introduced into the test vessel.
V dokončnem preskusu je končna točka plodnost (tj. število ustvarjenih mladičev).
Test chemical insoluble in water
Vendar je treba v končno poročilo, tako kot pri preskusu za določanje območja delovanja, zabeležiti vse druge škodljive znake.
For chemicals insoluble in water but soluble in organic solvents, the test chemical can be dissolved in the smallest possible volume of a suitable vehicle (e.g. acetone).
Za statistično analizo morata biti izračunana aritmetična sredina in standardni odklon za razmnoževanje pri posamezni tretirani skupini in kontroli.
Only volatile solvents should be used.
Če je izvedena analiza variance, je mogoče standardni odklon, s, in stopinje prostosti, df, nadomestiti z oceno skupne variance, pridobljene z analizo variance (ANOVA) oziroma z njenimi stopinjami prostosti, če varianca ni odvisna od koncentracije.
The vehicle is sprayed on or mixed with a small amount, for example 2,5 g, of fine quartz sand.
V tem primeru uporabite enosmerne analize variance kontrol in tretiranih skupin.
The vehicle is eliminated by evaporation under a fume hood for at least one hour.
Te vrednosti se navadno izračunajo s komercialno programsko opremo za statistiko, pri čemer so rezultati glede na posodo uporabljeni kot ponovitve.
This mixture of quartz sand and test chemical is added to the pre-moistened soil and thoroughly mixed after adding an appropriate amount of de-ionised water to obtain the moisture required.
Če se zdi, da je združevanje podatkov za negativne kontrole in kontrole s topilom bolj smiselno od preskušanja glede na enega od njih, jih je treba preskusiti, da se preveri, da med njimi ni značilnih razlik (za ustrezne preskuse glej odstavek 45 in Dodatek 7).
The final mixture is introduced into the test vessels.
Nadaljnje statistično preskušanje in sklepanje je odvisno od tega, ali so vrednosti ponovitev normalno porazdeljene in homogene glede na svojo varianco.
For chemicals that are poorly soluble in water and organic solvents, the equivalent of 2,5 g of finely ground quartz sand per test vessel is mixed with the quantity of test chemical to obtain the desired test concentration.
Ocena vrednosti NOEC
This mixture of quartz sand and test chemical is added to the pre-moistened soil and thoroughly mixed after adding an appropriate amount of de-ionised water to obtain the required moisture content.
Prednost mora imeti uporaba učinkovitih preskusov.
The final mixture is divided between the test vessels.
Uporabiti je treba informacije npr. iz predhodnih izkušenj s krožnim preskušanjem ali druge podatke iz preteklih študij o tem, ali so podatki približno normalno porazdeljeni.
The procedure is repeated for each test concentration and an appropriate control is also prepared.
Homogenost variance (homoskedastičnost) je bolj kritična.
Chemicals should not normally be tested at concentrations higher than 1000 mg/kg dry mass of soil.
Izkušnje kažejo, da varianca pogosto narašča z naraščajočo srednjo vrednostjo.
Testing at higher concentrations may however be required in accordance with the objectives of a specific test.
V teh primerih transformacija podatkov lahko privede do homoskedastičnosti.
PERFORMANCE OF THE TESTS
Vendar mora taka transformacija temeljiti na izkušnjah s podatki iz preteklih študij in ne na podatkih, ki se proučujejo.
Test groups and controls
Pri homogenih podatkih je treba izvesti več t-preskusov, kot je Williamsov preskus (α = 0,05, enostranski) (24)(25) ali, v nekaterih primerih, Dunnettov preskus (26)(27).
For each test concentration, an amount of test soil corresponding to 20 g dry weight is placed into the test vessel (see paragraphs 19-21).
Velja pripomniti, da je treba v primeru neenake ponovitve popraviti vrednosti t iz preglednice tako, kot sta predlagala Dunnett in Williams.
Controls, without the test chemical, are also prepared.
Včasih se odzivi zaradi velike variacije ne povečajo/zmanjšajo pravilno.
Food is added to each vessel in accordance with procedures described in paragraph 29.
V tem primeru velikega odklona od monotonosti je bolj primeren Dunnettov preskus.
Ten worms are randomly allocated to each test vessel.
Če obstajajo odkloni od homoskedastičnosti, je morda smiselno natančneje proučiti možne učinke na variance, da se ugotovi, ali je mogoče uporabiti t-preskuse brez prevelike izgube učinkovitosti (28).
The worms are carefully transferred into each test vessel and placed on the surface of the soil using, for example, jeweller's tweezers, hooks or loops.
Druga možnost je uporaba večkratnega U-preskusa, npr. Bonferronijevega U-preskusa po Holmu (29), ali – če ti podatki kažejo heteroskedastičnost, sicer pa so skladni z osnovnim monotonim razmerjem odmerek-odziv – drugega neparametričnega preskusa (npr. preskusa po Jonckheere-Terpstraju (30)(31) ali Shirleyjevega preskusa (32)(33)); uporaba takšnega preskusa ima v splošnem prednost pred t-preskusi za neenake variance.
The number of replicates for test concentrations and for controls depends on the test design used (see paragraph 34).
(Glej tudi shemo v Dodatku 7.)
The test vessels are positioned randomly in the test incubator and these positions are re-randomised weekly.
Če je izveden mejni preskus in so izpolnjeni predpogoji za postopke parametričnih preskusov (normalnost, homogenost), je mogoče uporabiti parni Studentov t-preskus ali, namesto tega, postopek Mann-Whitneyjevega U-preskusa (29).
If a vehicle is used for application of the test chemical, one control series containing quartz sand sprayed or mixed with solvent should be run in addition to the test series.
Ocena vrednosti ECx
The solvent or dispersant concentration should be the same as that used in the test vessels containing the test chemical.
Za izračun katere koli vrednosti ECx se po pridobitvi ustrezne funkcije odmerek-odziv za regresijsko analizo (linearno ali nelinearno) uporabijo srednje vrednosti glede na tretirano skupino.
A control series containing additional quartz sand (2,5 g per vessel) should be run for chemicals requiring administration in accordance with the procedures described in paragraph 21.
Za rast črvov v obliki zveznega odziva je mogoče vrednosti ECx oceniti z ustrezno regresijsko analizo (35).
The test temperature is 20 ± 2 °C.
Med primernimi funkcijami za kvantne podatke (smrtnost/preživetje in število ustvarjenih potomcev) so normalna sigmoidna funkcija in logistične ali Weibullove funkcije, ki vsebujejo od dva do štiri parametre, pri čemer nekatere lahko modelirajo tudi odzive v obliki hormeze.
To discourage worms from escaping from the soil, the test is carried out under controlled light-dark cycles (preferably 16 hours light and 8 hours dark) with illumination of 400 to 800 lux in the area of the test vessels.
Če je bila funkcija odmerek-odziv prilagojena z linearno regresijsko analizo, je treba v regresijski analizi pred ocenjevanjem vrednosti ECx najti značilni r2 (determinacijski koeficient) in/ali naklon, tako da se v enačbo, podano z regresijsko analizo, vstavi vrednost, ki ustreza x % srednje vrednosti kontrole.
In order to check the soil humidity, the vessels are weighed at the beginning of the test and thereafter once a week.
V skladu s Fiellerjem (navedeno v Finney (21)) ali z drugimi ustreznimi sodobnimi metodami se izračunajo 95-odstotne meje zaupanja.
Weight loss is replenished by the addition of an appropriate amount of deionised water.
Namesto tega se odziv lahko modelira kot odstotek ali razmerje parametra modela, ki se razlaga kot srednja vrednost odziva kontrole.
It should be noted that loss of water can be reduced by maintaining a high air-humidity (> 80 %) in the test incubator.
V teh primerih je mogoče običajno (logistično, Weibullovo) sigmoidno krivuljo pogosto enostavno prilagoditi rezultatom s postopkom regresije probit (21).
The moisture content and the pH, should be measured at the beginning and the end of both the range-finding test and the definitive test.
V teh primerih je treba utežno funkcijo prilagoditi za odzive metrik, kot je podano v Christensen (36).
Measurements should be made in control and treated (all concentrations) soil samples prepared and maintained in the same way as the test cultures but not containing worms.
Če pa je ugotovljena hormeza, mora analizo probit nadomestiti logistična ali Weibullova funkcija s štirimi parametri, prilagojena s postopkom nelinearne regresije (36).
Food should only be added to these soil samples at the start of the test to facilitate microbial activity.
Če podatkom ni mogoče prilagoditi primerne funkcije odmerek-odziv, je mogoče za oceno vrednosti ECx in njenih meja zaupanja uporabiti alternativne metode, kot so drseča povprečja po Thompsonu (22) in postopek prilagojene metode Spearman-Karber (23).
The amount of food added should be the same as that added to the test cultures.
Poročilo o preskusu mora vsebovati naslednje informacije:
It is not necessary to add further food to these vessels during the test.
fizikalno stanje in, kjer je ustrezno, fizikalno-kemijske lastnosti (npr. topnost v vodi, parni tlak),
Feeding
kemijski identifikacijski podatki preskusne kemikalije po nomenklaturi IUPAC, številka CAS, serija, skupina, strukturna formula in čistost,
A food capable of maintaining the enchytraeid population can be used.
datum izteka roka uporabnosti vzorca.
Rolled oats, preferably autoclaved before use to avoid microbial contamination (heating is also appropriate), have been found to be a suitable feeding material.
uporabljene preskusne živali:
Food is first provided by mixing 50 mg of ground rolled oats with the soil in each vessel before introducing the worms.
vrsta, znanstveno ime, vir organizmov in pogoji gojenja.
Thereafter, food is supplied weekly up to Day 21.
sestavine in priprava umetnih tal,
Feeding is not carried out on Day 28 since the adults have been removed at this stage and the juvenile worms need relatively little additional food beyond this point.
metoda uporabe preskusne kemikalije,
Feeding during the test comprises 25 mg of ground rolled oats per vessel placed carefully on the surface of the soil so as to avoid injuring the worms.
opis preskusnih pogojev, vključno s temperaturo, vsebnostjo vlage, vrednostjo pH itd.,
In order to reduce fungal growth, the oats flakes should be buried in the soil by covering with small amounts of soil.
celoten opis načrta in postopkov poskusa.
If food remains uneaten the ration should be reduced.
Rezultati preskusa:
Design for the range-finding test
smrtnost pri odraslih črvih po dveh tednih in število mladičev na koncu preskusa za določanje območja delovanja,
When necessary, a range-finding test is conducted with, for example, five test chemical concentrations of 0,1, 1,0, 10, 100, and 1000 mg/kg (dry weight of soil).
smrtnost pri odraslih črvih po treh tednih izpostavljenosti in celoten seznam mladičev na koncu dokončnega preskusa,
One replicate for each treatment and control is sufficient.
morebitni opaženi fizični ali patološki simptomi in spremembe vedenja pri preskusnih organizmih,
The duration of the range-finding test is two weeks.
vrednosti LC50, NOEC in/ali ECx (npr. EC50, EC10) za razmnoževanje, če je mogoče nekatere od njih uporabiti znotraj intervalov zaupanja, krivulja prilagojenega modela, uporabljena za njegov izračun, ter vse informacije in opažanja, ki pripomorejo k razlagi rezultatov.
At the end of the test, mortality of the worms is assessed.
Odstopanja od postopkov, opisanih v tej preskusni metodi, in morebitni nenavadni dogodki med preskusom.
A worm is recorded as dead if it has no reaction to a mechanical stimulus at the anterior end.
Poglavje C.8 te priloge, Strupenost za deževnike.
Additional information to mortality may also be useful in deciding on the range of concentrations to be used in the definitive test.
Part 1:
Changes in adult behaviour (e.g. the inability to dig into the soil; lying motionless against the glass wall of the test vessel) and morphology (e.g. the presence of open wounds) should therefore also be recorded along with the presence of any juveniles.
Part 2:
The latter can be determined using the staining method described in Appendix 6.
Determination of effects on reproduction, št. 11268-2.
The LC50 can be approximately determined by calculating the geometrical mean of mortality data.
Za to preskusno metodo se uporabljajo naslednje opredelitve:
In setting the concentration range for the definitive test, effects on the reproduction are assumed to be lower than the LC50 by a factor of up to 10.
V tem preskusu so učinkovite koncentracije izražene kot masa preskusne kemikalije na suho maso preskusnih tal.
However, this is an empirical relation ship and in any specific case it might be different.
(koncentracija brez smrtnega učinka) je koncentracija preskusne kemikalije, ki v določenem časovnem obdobju ne ubije nobenega od izpostavljenih preskusnih organizmov.
Additional observations made in the range-finding test such as the occurrence of juveniles can help refine the test chemical concentration range to be used for the definitive test.
V tem preskusu je LC0 izražena kot masa preskusne kemikalije na suho maso preskusnih tal.
In order for an accurate determination of the LC50 performing the test using at least four replicates each of the test chemical concentration and an adequate number of concentrations to cause at least four statistically significantly different mean responses at these concentrations) is recommended.
(srednja smrtna koncentracija) je koncentracija preskusne kemikalije, ki v določenem časovnem obdobju ubije 50 % izpostavljenih preskusnih organizmov.
A similar number of the concentrations and replicates for the controls are used when they are applicable.
V tem preskusu je LC50 izražena kot masa preskusne kemikalije na suho maso preskusnih tal.
Design for the definitive reproduction test
(popolnoma smrtna koncentracija) je koncentracija preskusne kemikalije, ki v določenem časovnem obdobju ubije 100 % izpostavljenih preskusnih organizmov.
Three designs are proposed based on recommendations arising from a ring test (2)
V tem preskusu je LC100 izražena kot masa preskusne kemikalije na suho maso preskusnih tal.
For determination of the NOEC, at least five concentrations in a geometric series should be tested.
LOEC (najnižja koncentracija z opaznim učinkom) je najnižja koncentracija preskusne kemikalije, ki ima statistično značilen učinek (p < 0,05).
Four replicates for each test concentration plus eight controls are recommended.
V tem preskusu je vrednost LOEC izražena kot masa preskusne kemikalije na suho maso preskusnih tal.
The concentrations should be spaced by a factor not exceeding 1,8.
Vse preskusne koncentracije nad vrednostjo LOEC morajo običajno kazati učinek, ki je statistično različen od kontrole.
For determination of the ECx (e.g. EC10, EC50), at least five concentrations should be tested and the concentrations should bracket ECx in order to enable ECx interpolation and not extrapolation At least four replicates for each test concentration and four control replicates are recommended.
Vsa odstopanja od zgoraj navedenega pri ugotavljanju vrednosti LOEC morajo biti utemeljena v poročilu o preskusu.
The spacing factor may vary, i.e. less than or equal to 1,8 in the expected effect range and above 1,8 at the higher and lower concentrations.
NOEC (koncentracija brez opaznega učinka) je najvišja koncentracija preskusne kemikalije tik pod vrednostjo LOEC, pri kateri ni opaznega učinka.
A combined approach allows for determination of both the NOEC and ECx.
Hitrost razmnoževanja je srednja vrednost števila mladih črvov, ki jih ustvari določeno število odraslih osebkov v preskusnem obdobju.
Eight treatment concentrations in a geometric series should be used.
Določanje največje zmogljivosti zadrževanja vode