natgillin/translations-raw huggingface dataset:
Finally, the nutrient solution is filled into 0,5 l Schott flasks and autoclaved at 121 °C for 20 minutes.
Koncentracije kemikalije i kontrolne skupine
The tenfold concentrated modified nutrient solution thus yielded can be kept sterile in a refrigerator (at 5-10 °C) for three months.
Koncentracije bi trebale slijediti geometrijski niz; faktor za koji se ispitne koncentracije razlikuju ne bi smio biti veći od 3,2.
MAINTENANCE OF STOCK CULTURE
Prethodna saznanja o toksičnosti ispitivane kemikalije na temelju ispitivanja za utvrđivanje raspona mogu pomoći u odabiru prikladnih ispitnih koncentracija.
In this Appendix 3 the stock culture of Myriophyllum spicatum L [1], a submersed aquatic dicotyledon, a species of the water milfoils family is described.
Da bi se pri određivanju ECx osigurala primjerena razina pouzdanosti, ECx mora biti između najviše i najniže ispitne koncentracije.
For sediment-free stock culture under laboratory conditions, sterile plants are required.
Upotrebom više ispitnih koncentracija poboljšat će se interval pouzdanosti za dobivenu vrijednost ECx.
Sterile plants are available from the ecotoxicology laboratory of the German Umweltbundesamt (Federal Environment Agency of Germany).
Za određivanje LOEC/NOEC (krajnja točka po izboru), najniža ispitna koncentracija mora biti dovoljno niska da se rast ne razlikuje značajno od rasta kontrolnih biljaka.
Alternatively, test organisms can be prepared from non-sterile plants in accordance with ASTM designation E 1913-04.
Isto tako, najviša ispitna koncentracija trebala bi biti dovoljno visoka da rast bude znatno niži od rasta kontrolnih biljaka.
See below — extracted from the ASTM Standard Guide — the procedure for culturing Myriophyllum sibiricum collected from field:
Upotrebom više ponovljenih uzoraka poboljšat će se statistička pouzdanost najviše koncentracije bez učinka / analize varijance (ANOVA).
Since the modified Andrews' medium contains sucrose (which stimulates the growth of fungi and bacteria), all material, solutions and culturing be conducted under sterile conditions.
Kad ispitivanje za utvrđivanje raspona ukazuje na to da ispitivana kemikalija nema štetno djelovanje u koncentraciji do 100 mg/l ili do granice svoje topljivosti u ispitnom mediju, odnosno, ako je riječ o formulaciji, do granice disperzivnosti, može se provesti ispitivanje granične koncentracije kako bi se mogli usporediti odgovori kontrolne skupine s odgovorom jedne tretirane skupine (100 mg/l ili koncentracija koja odgovara granici topljivosti, ili 1000 mg/kg suhog sedimenta).
In addition, all flasks, dishes, bowls etc and other equipment undergo flame treatment at the sterile workbench just prior to use.
To ispitivanje treba provesti u skladu s općim načelima standardnog ispitivanja koncentracija-odgovor, s iznimkom da se savjetuje povećati minimalni broj ponovljenih uzoraka na šest ispitnih posuda po kontroli i koncentraciji.
Stock cultures can be maintained under reduced illumination and temperature (50 μE m– 2 s– 1, 20 ± 2 °C) for longer times without needing to be re-established.
Ispitne otopine obično se pripremaju razrjeđivanjem matične otopine, koja se priprema otapanjem ili disperzijom ispitivane kemikalije u Smartovu i Barkovu mediju s demineraliziranom (tj. destiliranom ili deioniziranom) vodom (vidjeti Dodatak 1.).
The Myriophyllum growth medium should be the same as that used for testing but other nutrient rich media can be used for stock cultures.
Najviša ispitna koncentracija ne bi smjela biti veća od topljivosti ispitivane kemikalije ili, ako je riječ o formulaciji, njezine disperzivnosti u ispitnim uvjetima.
The plant segments are distributed axenically over several 500 ml Erlenmeyer or/and 2000 ml Fernbach flasks, each filled with approximately 450 respectively 1000 ml modified Andrews' medium.
Za ispitivane kemikalije slabe topljivosti u vodi možda će biti potrebno pripremiti koncentriranu matičnu otopinu ili disperziju kemikalije upotrebljavajući organsko otapalo ili dispergent kako bi se olakšalo dodavanje točnih količina ispitivane kemikalije u ispitni medij te ubrzalo njezinu disperziju i otapanje.
Then, the flasks are axenically cellulose plug stoppered.
Treba svakako nastojati izbjeći upotrebu takvih otapala ili dispergenata.
In addition, thorough flame treatment of equipment at the sterile workbench just prior to use is absolutely necessary.
Neka od otapala u širokoj uporabi koja nemaju fitotoksično djelovanje u koncentracijama do 100 μl/l su aceton i dimetilformamid.
Dependent on number and size, the plants are to be transferred into fresh nutrient solution approximately every three weeks.
Ako se upotrebljava otapalo ili dispergent, treba navesti njegovu konačnu koncentraciju, koju treba svesti na najmanju moguću mjeru (≤ 100 μl/l).
Apices as well as segments of the stem middle part for this renewed culture can be used.
U tim uvjetima sve tretirane skupine i kontrole (s otapalom) trebale bi sadržavati istu koncentraciju otapala odnosno dispergenta.
Number and size of transferred plants (or segments of plants) are dependent on how many plants are needed.
Netretirani kontrolni ponovljeni uzorci koji ne sadržavaju otapalo ili dispergent također su uključeni u plan ispitivanja.
For example, you can transfer five shoot segments into one Fernbach flask and three shoot segments into one Erlenmeyer flask, each with a length of 5 cm.
Dodatne smjernice o upotrebi dispergenata dostupne su u Smjernicama OECD-a (11.).
Discard any rooted, flowering, dead or otherwise conspicuous parts.
Ispitni postupak varira s obzirom na način primjene ispitivane kemikalije (u vodenoj ili sedimentnoj fazi).
Cutting of plants for the stock and pre culture after 3 weeks of cultivation
Pri odabiru načina izlaganja u ispitivanju (statički postupak ili statički postupak s obnavljanjem, voda s dodatkom ili sediment s dodatkom) trebalo bi razmotriti očekivano ponašanje ispitivane kemikalije u sustavu voda-sediment.
waste
Sediment s dodatkom ponekad može imati prednost za kemikalije za koje se očekuje znatna razdioba u sedimentu.
pre culture
Faza uspostave kulture
stock culture
Od biljaka odabrane kulture odrežu se vrhovi zdravih izdanaka, tj. bez bočnih izdanaka, duljine 6 cm (± 1 cm).
Culturing of plants is to be performed in 500 ml Erlenmeyer and 2000 ml Fernbach flasks in a cooling incubator at 20 ± 2 °C with continuously light at approximately 100-150 μE m– 2 s– 1 or 6000-9000 Lux (emitted by chamber illumination with colour temperature “warm white light”).
Za plan ispitivanja A (jedan izdanak u loncu, tri lonca u posudi) u svaki lonac sadi se po jedan izdanak.
Culturing of plants in a cooling incubator with chamber illumination
Za plan ispitivanja B (tri izdanka u loncu, jedan lonac u posudi) u svaki lonac sa sedimentom sadi se po četiri do pet vrhova izdanka.
Chemically clean (acid-washed) and sterile glass culture vessels should be used and aseptic handling techniques employed.
U oba slučaja potrebno je posaditi izdanke u više lonaca kako bi se na početku ispitivanja mogle odabrati jednolike biljke te kako bi bilo rezervnih biljaka koje se mogu upotrijebiti za pregled rasta korijena neposredno prije tretiranja i onih koje se mogu ubrati radi mjerenja biomase i duljine izdanka 0. dan.
In the event of contamination of the stock culture e.g. by algae, fungi and/or bacteria a new culture should be prepared or a stock culture from another laboratory should be used to renewal of the one culture.
Izdanci se sade tako da približno 3 cm, na kojima se nalaze najmanje dva nodija, budu ispod površine sedimenta.
Carl von Linné (* May, 23th, 1707 in Råshult/Älmhult; † January, 10th, 1778 in Uppsala).
Lonci se zatim prenose u ispitne posude u istim okolnim uvjetima kao za fazu izlaganja te se sedam dana drže u Smartovu i Barkovu mediju kako bi se počeo razvijati korijen.
MAINTENANCE OF PRE-CULTURE AND PREPARATION OF TEST ORGANISM FOR TESTING
Nakon tog razdoblja nekoliko se biljaka u rezervnim loncima uzima za pregled rasta korijena.
To obtain pre-culture, cut shoots of stock culture into segments with two whorls each; put segments into Fernbach flasks filled with modified Andrews' medium (with 3 % sucrose).
Ako nije vidljiv rast korijena (tj. ne vide se vrhovi korijena), fazu uspostave kulture treba produžiti dok ne bude vidljiv rast korijena.
Each flask can contain up to 50 shoot segments.
Taj se korak preporučuje kako bi se osiguralo da biljke aktivno rastu u vrijeme započinjanja ispitivanja.
However, care is to be taken that the segments are vital and do not have any roots and lateral branches or their buds (see figure 1 in Appendix 3).
Odabir jednolikog biljnog materijala
The pre-culture organisms are cultured for 14 to 21 days under sterile conditions in an environmental chamber with alternating 16/8 hour light/dark phases.
Za plan ispitivanja A (jedan izdanak u loncu, tri lonca u posudi) jednoliki lonci biraju se prije započinjanja ispitivanja.
Light intensity selected from the range of 100-150 μE m– 2 s– 1.
Za plan ispitivanja B (tri izdanka u loncu, jedan lonac u posudi), suvišne se biljke uklanjaju kako bi ostale tri biljke jednake veličine i izgleda.
Since the modified Andrews' medium contains sucrose (which stimulates the growth of algae, fungi and bacteria), test chemical solutions should be prepared and culturing be conducted under sterile conditions.
Izlaganje u vodenoj fazi
Sterilisation is carried out via heated air treatment (210 °C) for 4 hours or autoclaving for 20 minutes at 121 °C.
Lonci s biljkama koji su odabrani zbog jednolikosti stavljaju se u ispitne posude kako je određeno u planu ispitivanja.
In addition, all flasks, dishes, bowls etc. and other equipment undergo flame treatment at the sterile workbench just prior to use.
Zatim se u ispitne posude dodaje Smartov i Barkov medij.
Shoots are axenically removed from the pre-culture flasks, choosing material that is as homogeneous as possible.
Treba paziti da se izbjegne remećenje sedimenta.
Each testing requires at least 60 test organisms (testing with eight test chemical concentrations).
Zato se mediji mogu dodavati koristeći se lijevkom ili plastičnim diskom za pokrivanje sedimenta dok se medij ulijeva u ispitne posude, uz uvjet da se disk ukloni odmah nakon toga.
For testing, take fresh lateral branches from pre-cultures, shorten them to 2,5 cm from base (measured with ruler) and transfer them into a beaker containing sterile modified Andrews' medium.
Biljni lonci mogu se staviti u ispitne posude i nakon dodavanja medija.
These fresh lateral branches can be used for the sediment-free Myriophyllum spicatum toxicity test.
U oba slučaja svježi mediji mogu se upotrebljavati na početku faze izlaganja, ako je potrebno, kako bi se smanjilo potencijalno gomilanje algi i bakterija ili omogućila priprema jedinstvenih šarži ispitne otopine među ponovljenim uzorcima.
Cutting of plants from the pre culture for the sediment-free Myriophyllum spicatum toxicity test
Prije ili nakon dodavanja medija mjeri se duljina izdanka iznad sedimenta.
waste or further cultivation
Odgovarajuća količina ispitivane kemikalije može se dodati u ispitni medij prije nego se on doda u ispitne posude.
fresh lateral branches for the test
Ispitivana kemikalija može se dodati u medij i nakon što je on dodan u ispitne posude.
C.51 water-sediment Myriophyllum spicatum toxicity test
U tom slučaju treba paziti da se ispitivana kemikalija jednoliko rasporedi u cijelom ispitnom sustavu bez remećenja sedimenta.
This test method is equivalent to the OECD test guideline 239 (2014).
U svakom slučaju na početku ispitivanja bilježi se izgled ispitnog medija (npr. jasan, mutan itd.).
Test methods are available for the floating, monocotyledonous aquatic plant, Lemna species (1) and for algal species (2).
Izlaganje u sedimentu
These methods are routinely used to generate data to address the risk of test chemicals, in particular chemicals with herbicidal activity, to non-target aquatic plant species.
Sedimenti s dodatkom odabrane koncentracije pripremaju se dodavanjem otopine ispitivane kemikalije izravno u svježi sediment.
However, in some cases, data for additional macrophyte species may be required.
Matična otopina ispitivane kemikalije otopljena u deioniziranoj vodi miješa se s formuliranim sedimentom valjkastim mlinom, miješalicom hrane ili ručnim miješanjem.
Recent guidance published from the Society of Environmental Toxicology and Chemistry (SETAC) workshop on Aquatic Macrophyte Risk Assessment for Pesticides (AMRAP) proposed that data for a rooted macrophyte species may be required for test chemicals where Lemna and algae are known not to be sensitive to the mode of action or if partitioning to sediment is a concern, leading to exposure via root uptake (3).
Ako je ispitivana kemikalija slabo topljiva u vodi, može se otopiti u što je moguće manjem volumenu prikladnog organskog otapala (npr. heksan, aceton ili kloroform).
Based on current understanding and experience, Myriophyllum spp were selected as the preferred species in cases where data are required for a submerged, rooted dicotyledonous species (4) (5) (6).
Ta se otopina potom miješa s otprilike 10 g finog kvarcnog pijeska po jednoj ispitnoj posudi.
The water-sediment Myriophyllum spicatum test method complements the sediment-free Myriophyllum spicatum Toxicity Test (7).
Otapalo se pusti da ispari, a pijesak se zatim miješa s prikladnom količinom sedimenta po ispitnoj laboratorijskoj čaši.
This document describes a test method, which allows assessment of the effects of a test chemical on the rooted, aquatic plant species Myriophyllum spicatum, growing in a water-sediment system.
Samo lako hlapljiva sredstva mogu se upotrebljavati za otapanje, dispergiranje ili emulgiranje ispitivane kemikalije.
The test method is based partly on existing methods (1) (2) (8) and takes account of recent research related to the risk assessment of aquatic plants (3).
Treba imati na umu da se u završnoj fazi pripreme sedimenta mora uzeti u obzir volumen/masa pijeska s dodatkom ispitivane kemikalije (sediment stoga treba pripremiti s manje pijeska).
The water-sediment method has been validated by an international ring-test conducted with Myriophyllum species grown under static conditions, which were exposed to the test chemical through applications made via the water column (9).
Treba paziti da ispitivana kemikalija koja se dodaje u sediment bude temeljito i ravnomjerno raspoređena u njemu.
However, the test system is readily adapted to allow for exposure via spiked sediment or exposure via the water phase in semi-static or pulsed-dose scenarios, although these scenarios have not been formally ring tested.
Sediment s dodatkom stavlja se u lonce (kako je opisano).
Furthermore, the general method can be used for other rooted, submerged and emergent species including other Myriophyllum species (e.g. Myriophyllum aquaticum) and Glyceria maxima (10).
Biljke odabrane zbog jednolikosti i odgovarajućeg sustava korijenja vade se iz lonaca kojima se koristilo u fazi uspostave kulture i presađuju u sediment s dodatkom kako je opisano.
Modifications of test conditions, design and duration may be required for alternative species.
Lonci s biljkama stavljaju se u ispitne posude kako je određeno u planu ispitivanja.
In particular, more work is needed to define appropriate procedures for Myriophyllum aquaticum.
Zatim se u ispitne posude dodaje Smartov i Barkov medij, oprezno (s pomoću lijevka) kako bi se izbjeglo remećenje sedimenta.
These options are not presented in detail in this test method, which describes the standard approach for exposure of Myriophyllum spicatum in a static system via the water phase.
Prije ili nakon dodavanja medija mjeri se duljina izdanka iznad sedimenta.
This test method applies to substances, for which the test method has been validated, (see details in the ring test report (9)) or to formulations or known mixtures.
Održavanje razine vode tijekom ispitivanja
A Myriophyllum test may be conducted to fulfil a Tier 1 data requirement triggered by potential test chemical partitioning to sediment or mode of action/selectivity issues.
Mora se zabilježiti konačni volumen vode i označiti razina vode na svakoj ispitnoj posudi.
Equally, a laboratory-based Myriophyllum test may be required as part of a higher-tier strategy to address concerns over the risk to aquatic plants.
Ako tijekom ispitivanja voda ispari za više od 10 %, trebalo bi nadomjestiti razinu vode destiliranom vodom.
The specific reason for conducting a test will determine the route of exposure (i.e. via water or sediment).
Ako je potrebno, laboratorijske se čaše mogu lagano pokriti prozirnim pokrovom, npr. prozirnim plastičnim poklopcima, kako bi se smanjilo isparavanje i kontaminacija sporama algi.
Before use of this test method for the testing of a mixture intended for a regulatory purpose, it should be considered whether, and if so why, it may provide adequate results for that purpose.
Upotrebljava se toplo ili hladno bijelo fluorescentno svjetlo za osvjetljenje od otprilike 140 (± 20) μE·m– 2 s– 1 izmjereno kao fotosintetski aktivno zračenje (400 – 700 nm) na površini vode te u ciklusu svjetlost-tama od 16: 8 h.
The test is designed to assess chemical-related effects on the vegetative growth of Myriophyllum plants growing in standardised media (water, sediment and nutrients).
Odstupanja od odabranog osvjetljenja u ispitnom prostoru ne bi smjela biti veća od ± 15 %.
For this purpose, shoot apices of healthy, non-flowering plants are potted in standardised, artificial sediment, which is supplemented with additional nutrients to ensure adequate plant growth, and then maintained in Smart and Barko medium (Appendix 1).
Temperatura u ispitnim posudama trebala bi biti 20 ± 2 °C.
After an establishment period to allow for root formation, plants are exposed to a series of test concentrations added to the water column.
pH-vrijednost kontrolnog medija tijekom ispitivanja ne bi se smjela povećati za više od 1,5 jedinica.
Alternatively, exposure via the sediment may be simulated by spiking the artificial sediment with the test chemical and transplanting plants into this spiked sediment.
Ipak, zbog odstupanja za više od 1,5 jedinica ne dovodi se u pitanje valjanost ispitivanja ako se može dokazati da su zadovoljeni navedeni kriteriji valjanosti.
To quantify chemical-related effects, growth in the test solutions is compared with the growth of the control plants, and the concentration causing a specified x % inhibition of growth is determined and expressed as the ECx; “x” can be any value depending on the regulatory requirements, e.g. EC10 EC20 and EC50.
Nakon faze uspostave kulture i neposredno prije tretiranja (tj.
Both average specific growth rate (estimated from assessments of shoot length, shoot fresh weight and shoot dry weight) and yield (estimated from the increase in shoot length, shoot fresh weight and shoot dry weight) of untreated and treated plants should be determined.
0. dan) rezervne biljke iz pet nasumično odabranih lonaca za plan s tri biljke u loncu ili 15 lonaca za plan s jednom biljkom u loncu izdvajaju se za procjenu duljine izdanka i svježe i suhe mase kako je opisano u nastavku.
Information on the test chemical which may be useful in establishing the test conditions includes the structural formula, composition in the case of multi-constituent substances, UVCBs, mixtures or formulations, purity, water solubility, stability in water and light, acid dissociation constant (pKa), partition coefficient octanol-water (Kow), if available Kd to sediments, vapour pressure and biodegradability.
vizualne procjene zdravlja biljke bilježe se najmanje tri puta tijekom razdoblja izlaganja (npr.
Water solubility and vapour pressure can be used to calculate Henry's Law constant, which will indicate whether significant losses of the test chemical during the test period are likely.
0., 7. i 14. dan),
If losses of the test chemicals are likely, the losses should be quantified and the subsequent steps to control such losses should be documented.
procjene svježe i suhe mase izdanaka provode se na kraju ispitivanja (14. dan).
Where information on the solubility and stability of the test chemical(s) is uncertain, it is recommended that these properties are assessed under the conditions of the test, i.e. growth medium, temperature, lighting regime to be used in the test.
Duljina izdanka mjeri se pomičnim mjerilom.
Note: when light dependent peroxidising herbicides are tested, the laboratory lighting used should contain the equivalent presence of solar ultraviolet light found in natural sunlight.
Ako ima bočnih izdanaka potrebno je utvrditi njihov broj i duljinu.
The pH should be measured and adjusted in the test medium as appropriate.
Pri vizualnim procjenama zdravlja biljaka bilježe se izgled biljaka i opće stanje ispitnog medija.
The pH control of the test medium is particularly important, e.g. when testing metals or chemicals which are hydrolytically unstable.
Zapažanja koja je potrebno zabilježiti uključuju:
Further guidance for testing chemicals with physical-chemical properties that make them difficult to test is provided in a OECD Guidance Document (11).
nekrozu, klorozu i druge promjene boje, npr. pretjerano crvenilo u odnosu na kontrolne biljke,
For the test results to be valid, the mean total shoot length and mean total shoot fresh weight in control plants at least double during the exposure phase of the test.
kontaminaciju bakterijama ili algama,
In addition, control plants must not show any visual symptoms of chlorosis and should be visibly free from contamination by other organisms such as algae and/or bacterial films on the plants, at the surface of the sediment and in the test medium.
anomalije rasta kao što su zaostajanje u rastu, promjene duljine između nodija, deformacije izdanaka/listova, umnožavanje bočnih izdanaka, gubitak listova, gubitak turgora i fragmentacija stabljike,
The mean coefficient of variation for yield based on measurements of shoot fresh weight (i.e. from test initiation to test termination) in the control cultures does not exceed 35 % between replicates.
zdravlje korijena procjenjuje se vizualno na kraju ispitivanja uz pažljivo ispiranje sedimenta s korijena kako bi se mogao promatrati sustav korijena.
A reference chemical(s), such as 3,5-dichlorophenol used in the ring test (9), should be periodically tested in order to check the performance of the test procedure over time.
U nastavku je prijedlog ljestvice za procjenu u odnosu na kontrolne biljke:
The ring test data indicate that the mean EC50 values of 3,5-DCP for the different response variables were between 4,7 and 6,1 mg/l (see the ring-test report for details of anticipated confidence interval around these values).
nema korijena
It is advisable to test a reference chemical at least twice a year or, where testing is carried out infrequently, in parallel with the definitive toxicity tests.
malo korijena
A guide to expected EC50 values for 3,5-DCP is provided in the Statistical Report of the International Ring-test (9).
srednje razvijen korijen
Test apparatus
vrlo dobro razvijen korijen, slično kontrolnim biljkama.
The test should be conducted under controlled environmental conditions, i.e. in a growth chamber, growth room or laboratory, with controllable day length, lighting and temperature (see section “Test conditions”, paragraphs 56-58).
Svježa masa procjenjuje se na početku i kraju ispitivanja; izdanak se odreže na razini sedimenta i suši upijajućim materijalom prije vaganja.
Stock cultures should be maintained separately from test vessels.
Treba paziti da se uklone čestice sedimenta koje mogu prionuti uz dno izdanka.
The study should be conducted using glass test vessels such as aquaria or beakers; 2-l glass beakers (approximately 24 cm high and 11 cm in diameter) are commonly used.
Izdanak se zatim stavi u sušionik na otprilike 60 °C i suši do konstantne mase, a zatim se ponovno važe i bilježi se suha masa.
However, other (i.e. larger) vessels may be suitable provided that there is sufficient depth of water to allow unlimited growth and keep the plants submerged throughout the test duration.
U tablici 1. prikazane su minimalne biološke procjene koje je potrebno provesti tijekom ispitivanja.
Plastic or glass plant pots (approximately 9 cm diameter and 8 cm high and 500 ml volume) may be used as containers for potting the plants into the sediment.
Plan procjena