natgillin/translations-raw huggingface dataset:
Any suitable water, natural water (surface or ground water), reconstituted water (see Appendix 2) or dechlorinated tap water are acceptable as culturing water and test water, if chironomids will survive in it for the duration of the culturing and testing without showing signs of stress.
Voda s primešano preskusno snovjo je preskusna voda, ki ji je bila dodana preskusna kemikalija.
At the start of the test, the pH of the test water should be between 6 and 9 and the total hardness not higher than 400 mg/l as CaCO3.
Priporočila za gojenje Chironomus riparius
However, if there is an interaction suspected between hardness ions and the test chemical, lower hardness water should be used (and thus, Elendt Medium M4 should not be used in this situation).
Ličinke Chironomus se lahko gojijo v kristalizirkah ali večjih posodah.
The same type of water should be used throughout the entire study.
Na dno posode se nanese tanka plast finega kremenovega peska (debela približno od 5 do 10 mm).
The water quality characteristics listed in Appendix 4 should be measured at least twice a year or when it is suspected that these characteristics may have changed significantly.
Za primeren substrat se je izkazal tudi kieselgur (npr. Merck, Art 8117) (zadošča tanjša plast, debela samo nekaj mm).
Stock solutions — Spiked water
Nato se doda primerna voda do globine več cm.
a.
Vodo je treba po potrebi dotočiti, da se nadomestijo izgube zaradi izhlapevanja in prepreči izsušitev.
Test concentrations are calculated on the basis of water column concentrations, i.e. the water overlying the sediment.
Voda se po potrebi lahko zamenja.
Test solutions of the chosen concentrations are usually prepared by dilution of a stock solution.
Zagotoviti je treba rahlo prezračevanje.
Stock solutions should preferably be prepared by dissolving the test chemical in test water.
Posode, v katerih se gojijo ličinke, morajo biti v primerni kletki, ki bo preprečila pobeg odraslih preobraženih osebkov.
The use of solvents or dispersants may be required in some cases in order to produce a suitably concentrated stock solution.
Kletka mora biti dovolj velika, da lahko preobraženi odrasli osebki rojijo, sicer se lahko zgodi, da ne bo parjenja (najmanjša velikost je približno 30 × 30 × 30 cm).
Examples of suitable solvents are acetone, ethylene glycol monoethyl ether, ethylene glycol dimethylether, dimethylformamide and triethylene glycol.
Kletke morajo biti v prostoru s sobno temperaturo ali prostoru s konstantnim ozračjem pri 20 ± 2 °C s 16-urnim obdobjem osvetljenosti (jakost približno 1000 luksov) in 8 urami teme.
Dispersants which may be used are Cremophor RH40, Tween 80, methylcellulose 0,01 % and HCO-40.
Po poročilih lahko manj kot 60-odstotna relativna vlažnost zraka ovira razmnoževanje.
The solubilising agent concentration in the final test medium should be minimal (i.e. ≤ 0,1 ml/l) and should be the same in all treatments.
Uporabi se lahko vsaka primerna naravna ali sintetična voda.
When a solubilising agent is used, it should have no significant effects on survival as revealed by a solvent control in comparison with a negative (water) control.
Običajno se uporabljajo voda iz vodnjaka, deklorirana vodovodna voda in umetni mediji (npr. medij Elendt ‚M4‘ ali ‚M7‘, glej v nadaljevanju).
However, every effort should be made to avoid the use of such materials.
Vodo je treba pred uporabo prezračevati.
Stock solutions — Spiked sediment
Voda za gojenje se lahko po potrebi obnovi, tako da se uporabljena voda iz posod za gojenje skrbno pretoči ali izsesa, ne da bi se pri tem uničil ovoj ličink.
b.
Hranjenje ličink
Spiked sediments of the chosen concentration are usually prepared by addition of a solution of the test chemical directly to the sediment.
Ličinke Chironomus je treba hraniti s približno 250 mg ribje hrane v kosmičih (Tetra Min®, Tetra Phyll® ali druga podobna zaščitena znamka ribje hrane) na posodo na dan.
A stock solution of the test chemical dissolved in deionised water is mixed with the formulated sediment by rolling mill, feed mixer or hand mixing.
Hrana se lahko daje v obliki suhega mletega prahu ali kot suspenzija v vodi:
If poorly soluble in water, the test chemical can be dissolved in as small a volume as possible of a suitable organic solvent (e.g. hexane, acetone or chloroform).
1,0 g hrane v kosmičih se doda 20 ml vode za redčenje in zmeša, da nastane homogena mešanica.
This solution is then mixed with 10 g of fine quartz sand for each test vessel.
Ta pripravek se lahko daje v količini približno 5 ml na posodo na dan (pred uporabo ga je treba pretresti).
The solvent is allowed to evaporate and it should be totally removed from sand;
Starejše ličinke lahko dobijo več hrane.
the sand is then mixed with the suitable amount of sediment.
Hranjenje se prilagaja kakovosti vode.
Only agents which volatilise readily can be used to solubilise, disperse or emulsify the test chemical.
Če gojišče postane ‚moteno‘, je treba hranjenje zmanjšati.
It should be born in mind that the sand provided by the test chemical and sand mixture, should be taken into account when preparing the sediment (i.e. the sediment should thus be prepared with less sand).
Dodajanje hrane je treba skrbno spremljati.
If necessary, subsamples can be analysed to determine degree of homogeneity.
Premalo hrane bo povzročilo selitev ličink v vodni stolpec, preveč hrane pa povečano mikrobno dejavnost in zmanjšane koncentracije kisika.
The test design relates to the selection of the number and spacing of the test concentrations, the number of vessels at each concentration, the number of larvae per vessel, the number of crystallising dishes and breeding cages.
Oboje lahko privede do počasnejše rasti.
Designs for ECx, NOEC and a limit test are described below.
Ko se pripravijo nove posode za gojenje, se lahko dodajo tudi celice nekaterih zelenih alg (npr. Scenedesmus subspicatus, Chlorella vulgaris).
Design for analysis by regression
Hranjenje preobraženih odraslih osebkov
The effect concentration (ECx) and the concentration range over which the effect of the test chemical is of interest, should be spanned by the test, such that the endpoint is not extrapolated outside the bounds of the data generated.
Nekateri raziskovalci so predlagali, da se lahko kot hrana za odrasle preobražene osebke uporabi vatna blazinica, namočena v nasičeno raztopino saharoze.
Extrapolation much below the lowest or above the highest concentration should be avoided.
Pri 20 ± 2 °C se bo preobrazba v odrasle osebke iz posod za gojenje ličink začela po približno 13 do 15 dneh.
A preliminary range-finding test according to Test Methods C.27 or C.28 may be helpful for selecting a suitable range of test concentrations.
Samce je mogoče preprosto razlikovati po pahljačastih tipalkah in suhem telesu.
For an ECx approach, at least five concentrations and eight replicates for each concentration are required.
Jajčna legla
For each concentration two breeding cages should be used (A and B).
Ko so v kletki za razmnoževanje odrasli osebki, je treba vse posode za gojenje ličink trikrat na teden preveriti glede morebitnih odloženih želatinastih jajčnih legel.
The eight replicates are divided into two groups of four replicates to serve each breeding cage.
Če so prisotna, jih je treba previdno odstraniti.
This merger of replicates is necessary due to the number of midges needed in the cage for sound reproduction assessments.
Prenesti jih je treba v majhno posodo, ki vsebuje vzorec vode za razmnoževanje.
However, the 2nd generation has eight replicates again, which are initiated from the exposed populations in the breeding cages.
Jajčna legla se uporabijo za pripravo nove posode za gojenje (npr. 2 do 4 jajčna legla/posodo) ali za preskuse strupenosti.
The factor between concentrations should not be greater than two (an exception could be made in cases when the dose response curve has a shallow slope).
Ličinke prvega stadija bi se morale izvaliti po 2 ali 3 dneh.
The number of replicates at each treatment can be reduced to six (three for each breeding case) if the number of test concentrations with different responses is increased.
Priprava novih posod za gojenje
Increasing the number of replicates or reducing the size of the test concentration intervals tends to lead to narrower confidence intervals around the ECX.
Ko so gojišča vzpostavljena, bi moralo biti mogoče novo posodo za gojenje za ličinke pripraviti vsak teden ali manj pogosto, odvisno od zahtev preskušanja, stare posode pa odstraniti, ko se ličinke preobrazijo v odrasle osebke.
Design for estimation of a NOEC
S takim sistemom bo najpreprosteje zagotovljena redna oskrba z odraslimi osebki.
For a NOEC approach, five test concentrations with at least eight replicates (4 for each breeding cage, A and B) should be used and the factor between concentrations should not be greater than two.
Končne koncentracije v preskusnih raztopinah ‚M4‘ in ‚M7‘
The number of replicates should be sufficient to ensure adequate statistical power to detect a 20 % difference from the control at the 5 % level of significance (α = 0,05).
Elendt (1990) je opisal medij ‚M4‘.
For the development rate, fecundity and fertility an analysis of variance (ANOVA) is usually appropriate, followed by Dunnett's test or Williams' test (22-25).
Medij ‚M7‘ se pripravi kot medij ‚M4‘, razen za snovi, navedene v tabeli 1, za katere so koncentracije pri ‚M7‘ štirikrat nižje kot pri ‚M4‘.
For the emergence ratio and sex ratio the Cochran-Armitage, Fisher's exact (with Bonferroni correction), or Mantel-Haentzal tests may be appropriate.
Preskusna raztopina se ne sme pripraviti po Elendt in Bias (1990), saj koncentracije NaSiO3 · 5H2O, NaNO3, KH2PO4 in K2HPO4, navedene za pripravo založnih raztopin, niso ustrezne.
A limit test may be performed (one test concentration and control(s)) if no effects are observed in the optional preliminary range-finding test up to a maximum concentration.
Priprava medija ‚M7‘
The purpose of the limit test is to indicate that any toxic effects of the test chemical are found at levels greater than the limit concentration tested.
Vsaka založna raztopina (I) se pripravi posebej, sestavljena založna raztopina (II) pa se pripravi iz teh založnih raztopin (I) (glej tabelo 1).
For water, 100 mg/l and for sediment 1000 mg/kg (dry weight) are suggested.
V 50 ml sestavljene založne raztopine (II), ki ji je dodana količina vsake založne raztopine makrohranilnih snovi, navedena v tabeli 2, se dolije deionizirana voda do 1 litra, da se pripravi medij ‚M7‘.
Usually, at least eight replicates for both the treatment and control are necessary.
Založna raztopina vitaminov se pripravi tako, da se trije vitamini dodajo v deionizirano vodo, kot je navedeno v tabeli 3, 0,1 ml sestavljene založne raztopine vitaminov pa se doda končnemu mediju ‚M7‘ malo pred uporabo.
Adequate statistical power to detect a 20 % difference from the control at the 5 % level of significance (α = 0,05) should be demonstrated.
Založna raztopina vitaminov se hrani zamrznjena v majhnih alikvotih.
With metric responses (e.g. development rate), the t-test is a suitable statistical method if data meet the requirements of this test (normality, homogeneous variances).
Medij se prezračuje in stabilizira.
An unequal-variance t-test or a non-parametric test, such as the Wilcoxon-Mann-Whitney test may be used, if these requirements are not fulfilled.
Založne raztopine elementov v sledeh za medija M4 in M7
With the emergence ratio, Fisher's exact test is appropriate.
Založne raztopine (I)
Preparation of the water-sediment system (water spiking)
Količina (mg) za pripravo 1 litra z deionizirano vodo
a.
Za pripravo sestavljene založne raztopine (II):
Formulated sediment (see paragraphs 13-14 and Appendix 3) is added to each test vessel and crystallising dish to form a layer of at least 1,5 cm (for the crystallising dish it may be somewhat lower) but maximally 3 cm.
zmešajo se naslednje količine (ml) založnih raztopin (I) in dopolnijo do 1 litra z deionizirano vodo
Water (see paragraph 15) is added so that the ratio of the depth of the sediment layer and the depth of the water does not exceed 1:4.
Končne koncentracije v preskusnih raztopinah (mg/l)
After preparation of the test vessels the sediment-water system should be left under gentle aeration for approximately seven days prior to addition of the first instar larvae of the 1st or 2nd generation (see paragraph 14 and Appendix 3).
MnCl2 · 4H2O [1]
The sediment-water system of the crystallising dishes is not aerated during the test, since they do not need to support larval survival (before hatching the egg ropes are already collected).
SrCl2 · 6H2O [1]
To avoid separation of sediment ingredients and re-suspension of fine material during addition of test water in the water column, the sediment can be covered with a plastic disc while water is poured onto it.
MoO4 · 2H2O [1]
The disc is removed immediately afterwards.
CuCl2 · 2H2O [1]
Other devices may also be appropriate.
CaCl2 · 6H2O
Preparation of the water-sediment system (spiked sediment)
EDTA · 2H2O [1] [2]
b.
FeSO4 · 7H2O [1] [2]
The spiked sediments prepared according to paragraph 16b are placed in the vessels and crystallising dish and overlying water is added to produce a sediment-water volume ratio of 1:4.
Založne raztopine makrohranilnih snovi za medija M4 in M7
The depth of the sediment layer should be in the range of 1,5 to 3 cm (it may be somewhat lower for the crystallising dish).
Količina za pripravo 1 litra z deionizirano vodo
To avoid separation of sediment ingredients and re-suspension of fine material during addition of test water in the water column, the sediment can be covered with a plastic disc while water is poured onto it, and the disc removed immediately afterwards.
Količina založnih raztopin makrohranilnih snovi za pripravo medijev M4 in M7
Appropriate equilibration time is sediment and chemical specific, and can be in the order of hours to days and in rare cases up to five weeks.
Založna raztopina vitaminov za medija M4 in M7
As this would leave time for degradation of many chemicals, equilibrium is not awaited but an equilibration period of 48 hours is recommended.
Vse tri raztopine vitaminov so združene v eno.
However, when the degradation half-life of the chemical in sediment is known to be long (see paragraph 8), the equilibration time may be extended.
Količina dodane založne raztopine vitaminov za pripravo medijev M4 in M7
At the end of this further equilibration period, the concentration of the test chemical should be measured in the overlying water, the pore water and the sediment, at least at the highest concentration and a lower one (see paragraph 38).
Tiamin hidroklorid
These analytical determinations of the test chemical allow for calculation of a mass balance and expression of results based on measured concentrations.
Cianokobalamin (B12)
Test vessels should be covered (e.g. by glass plates).
Biotin
If necessary, during the study the water levels may be topped up to the original volume in order to compensate for evaporation.
Te snovi se pri M4 in M7 razlikujejo, kot je navedeno zgoraj.
This should be performed using distilled or deionised water to prevent any build-up of salts.
Te raztopine se pripravijo posamezno, nato zlijejo skupaj in takoj avtoklavirajo.
Crystallising dishes in the breeding cages are not covered and may, but do not need to be adjusted to compensate for water loss during the test period, since the egg ropes are only in contact with the water for about one day and the dishes are only used during a short phase of the test.
Priprava formulirane usedline
Addition of test organisms
SESTAVA USEDLINE
Four to five days before adding the first instar larvae for the 1st generation, egg masses should be taken from the culture and placed in small vessels in culture medium.
Sestava formulirane usedline mora biti naslednja:
Aged medium from the stock culture or freshly prepared medium may be used.
šotni mah, čim bližje pH 5,5–6,0, brez vidnih ostankov rastlin, fino mlet (velikost delcev ≤ 1 mm) in sušen na zraku
In any case, a small amount of food, e.g. a few droplets of filtrate from a finely ground suspension of flaked fish food, should be added to the culture medium (see Appendix 2).
4–5
Only freshly laid egg masses should be used.
> 50 % delcev mora biti velikih 50–200 μm
Normally, the larvae begin to hatch a couple of days after the eggs are laid (2 to 3 days for C. riparius at 20 °C and 1 to 4 days for C. dilutus at 23 °C and C. yoshimatsui at 25 °C) and larval growth occurs in four instars, each of 4-8 days duration.
CaCO3, zdrobljen v prah, kemijsko čist.
First instar larvae (maximum 48 h post hatching) should be used in the test.
0,05–0,1
The instar stage of larvae can potentially be checked using head capsule width (7).
prevodnost ≤ 10 μS/cm
Twenty first instar larvae for the 1st generation are allocated randomly to each test vessel containing the sediment-water system, using a blunt pipette.
PRIPRAVA
In the spiked sediment design, exposure starts with the addition of the larvae.
Priporoča se, da se formulirana usedlina pred uporabo v preskusu strupenosti s trzačami sedem dni kondicionira v enakih pogojih, kot vladajo v poznejšem preskusu.
Spiking the overlying water
SHRANJEVANJE
Twenty-four hours after adding the first instar larvae for the 1st generation, the test chemical is spiked into the overlying water column, and slight aeration is again supplied (for possible amendments of the test design, see paragraph 7).
Suhe sestavine za pripravo umetne usedline se lahko shranjujejo v suhem in hladnem prostoru pri sobni temperaturi.
Small volumes of the test chemical stock solutions are applied below the surface of the water using a pipette.
Formulirana (mokra) usedlina se pred uporabo v preskusu ne sme shranjevati.
The overlying water should then be mixed with care not to disturb the sediment.
Uporabiti jo je treba takoj po 7-dnevnem obdobju kondicioniranja, s katerim se njena priprava konča.
In the spiked water design, exposure starts with the spiking of the water (i.e. one day after addition of the larvae).
OECD (1984).
Collecting emerged adults
Earthworm, Acute Toxicity Test, Test Guideline No.
Emerged midges of the 1st generation are collected at least once, but preferably twice a day (see point 36) from the test vessels using an aspirator, exhauster or similar device (see Appendix 5).
207, Guidelines for the Testing of Chemicals, OECD, Pariz.
Special care should be taken not to damage the adults.
10–20.
The collected midges from four test vessels within one treatment are released into a breeding cage to which they had been previously assigned.
Kemijske značilnosti sprejemljive vode za redčenje
At the day of first (male) emergence, crystallising dishes are spiked by pipetting a small volume of the test chemical stock solution below the water surface (spiked water design).
SESTAVINA
The concentration of test chemical in the crystallising dish is nominally the same as in the treatment vessels which are assigned to that specific breeding cage.
KONCENTRACIJE
For the spiked sediment design, the crystallising dishes are prepared at around day 11 after the start of the exposure (i.e. addition of the 1st generation larvae) so that they can equilibrate for about 48 hours before the first egg ropes are produced.
trdota kot CaCO3
Egg ropes are collected from the crystallising dish in the breeding cage using tweezers or a blunt pipette.
Če se domneva, da bo prišlo do interakcije med ioni, ki povzročajo trdoto vode, in preskusno kemikalijo, je treba uporabiti vodo z manjšo trdoto (v takem primeru se torej ne sme uporabiti medij Elendt M4).
Each egg rope is placed into a vessel containing culture medium from the crystallising dish it was collected from (e.g. a well of a 12-well micro-plate together with at least 2,5 ml of medium).
Smernica za izvedbo preskusa
a.
B stekleni pokrov za preprečevanje pobega preobraženih trzač;
In the spiked water design, the sediment-water system for the 2nd generation is prepared by spiking the test chemical into the overlying water column ca.
C gladina vode;
1 hour before adding the first instar larvae to the test vessels.
D preskusna posoda (steklena časa s prostornino najmanj 600 ml);
Small volumes of the test chemical solutions are applied below the surface of the water using a pipette.
E plast usedline.
After spiking, slight aeration is supplied.
Primer sesalnika za zajetje odraslih trzač (puščici označujeta smer zračnega toka):
b.
Znotraj je odprtina steklene cevi (A) prekrita z bombažem in gazo (velikost mreže je približno 1 mm), da se preprečijo poškodbe trzač, ko jih sesalnik posesa;
In the spiked sediment design, the exposure vessels containing the sediment-water system for the 2nd generation are prepared in the same way as for the 1st generation.
C prozoren vsebnik (plastičen ali steklen, dolg približno 15 cm) za ujete trzače;
Twenty first instar larvae (maximum 48 h post hatching) of the 2nd generation are allocated randomly to each test vessel containing the spiked sediment-water system, using a blunt pipette.
D zamašek iz gume, preluknjan s cevjo (E).
Aeration of the water should be stopped while adding the first instar larvae to the test vessels and remain so for another 24 hours after addition of the larvae.
Zamašek D se odstrani z vsebnika C, da se trzače spustijo v kletko za razmnoževanje;
It is necessary to feed the larvae in the test vessels, preferably daily or at least three times per week.
Shematski prikaz preskusa življenjskega cikla:
Fish-food (a suspension in water or finely ground food, e.g. Tetra-Min or Tetra-Phyll; see details in Appendix 2) of 0,25 - 0,5 mg (0,35 - 0,5 mg for C. yoshimatsui) per larvae per day is an adequate amount of food for young larvae during the first 10 days of their development.
A prva generacija – preskusne posode, ki vsebujejo sistem z vodo in usedlino, osem ponovitev, 20 ličink prvega stadija na posodo;
Slightly more food may be necessary for older larvae:
B štiri preskusne posode za vsako kletko za razmnoževanje, A in B;
0,5 - 1,0 mg per larvae per day should be sufficient for the rest of the test.
C kletki za razmnoževanje (A in B) za rojenje, parjenje in odlaganje jajčec;
The food ration should be reduced in all treatments and control if fungal growth is seen or if mortality is observed in controls.
D kristalizirki za odlaganje nizov jajčec;
If fungal development cannot be stopped the test should be repeated.
E mikroplošči, ena vdolbinica za vsak niz jajčec;
The toxicological relevance of exposure via ingestion is generally higher in chemicals with a high affinity for organic carbon or chemicals covalently binding to the sediment.
F druga generacija – preskusne posode, ki vsebujejo sistem z vodo in usedlino, osem ponovitev, 20 ličink prvega stadija na posodo.
Hence, when testing chemicals with such properties, the amount of food necessary to ensure survival and natural growth of the larvae may be added to the formulated sediment before the stabilisation period, depending on the regulatory demand.
Ta preskusna metoda ustreza Smernici za preskušanje OECD (TG) 234 (2011).
To prevent deterioration of the water quality, plant material should be used instead of fish food, e.g. addition of 0,5 % (dry weight) finely ground leaves of stinging nettle (Urtica dioica), mulberry (Morus alba), white clover (Trifolium repens), spinach (Spinacia oleracea) or other plant material (Cerophyl or α-cellulose).
Temelji na sklepu iz leta 1998, da se pripravijo nove ali posodobijo obstoječe preskusne metode za spremljanje in preskušanje potencialnih povzročiteljev endokrinih motenj.
Addition of the complete ration of an organic food source to the sediment before spiking is not trivial with respect to water quality and biological performance (21), nor a standardised method, but recent studies provide indications that this method works (19) (26).
Preskus spolnega razvoja rib (FSDT) je bil opredeljen kot obetavna preskusna metoda, ki zajema občutljivo življenjsko fazo rib, ki se odzivajo na kemikalije, podobne estrogenu in androgenu.