natgillin/translations-raw huggingface dataset:
Illumination and temperature should be controllable and maintained at a constant level.
Taj bi utjecaj ostao neotkriven kad bi se računala samo duljina glavnog izdanka.
Test organism
Prosječna specifična brzina rasta za određeno razdoblje izračunava se kao logaritamsko povećanje varijabli rasta – duljine glavnog izdanka i još triju mjernih varijabli (ukupna duljina izdanka, svježa masa, suha masa ili broj pršljena) – primjenom formule u nastavku za svaki ponovljeni uzorak kontrolne i tretirane skupine:
Myriophyllum spicatum — a submersed aquatic dicotyledon — is a species of the water milfoils family.
prosječna specifična brzina rasta od vremena i do vremena j
Between June and August, inconspicuous pink-white flowers protrude above the water surface.
mjerna varijabla u ispitnoj ili kontrolnoj posudi u vremenu i
The plants are rooted in the ground by a system of robust rhizomes and can be found in the entire northern hemisphere in eutrophic, however non-polluted and more calciferous still waters with muddy substrate.
mjerna varijabla u ispitnoj ili kontrolnoj posudi u vremenu j
Myriophyllum spicatum prefers fresh water, but is found in brackish water as well.
t vremensko razdoblje od i do j
For the sediment-free toxicity test, sterile plants are required.
Za svaku tretiranu i kontrolnu skupinu treba izračunati srednju vrijednost brzine rasta i procjene varijance.
If the testing laboratory does not have regular cultures of Myriophyllum spicatum, sterile plant material may be obtained from another laboratory or (unsterile) plant material might be taken from the field or provided by a commercial supplier; if plants come from the field a taxonomic verification of the species should be envisaged.
Prosječnu specifičnu brzinu rasta treba izračunati za čitavo razdoblje ispitivanja (vrijeme ‚i’ u navedenoj formuli označava početak ispitivanja, a vrijeme ‚j’ kraj ispitivanja).
If collected from the field or provided by a commercial supplier, plants should be sterilised (1) and maintained in culture in the same medium as used for testing for a minimum of eight weeks prior to use.
Za svaku ispitnu koncentraciju i kontrolu treba izračunati srednju vrijednost prosječne specifične brzine rasta te procjene varijance.
Field sites used for collecting starting cultures have to be free of obvious sources of contamination.
Usto je potrebno procijeniti i brzinu rasta u pojedinim etapama tijekom razdoblja izlaganja kako bi se ocijenili učinci ispitivane kemikalije koji se javljaju (npr. pregledom logaritamski transformiranih krivulja rasta).
Great care should be taken to ensure that the correct species is obtained when collecting Myriophyllum spicatum from the field, especially in regions where it can hybridise with other Myriophyllum species.
Postotak inhibicije brzine rasta (Ir) tada se može izračunati za svaku ispitnu koncentraciju (tretiranu skupinu) prema sljedećoj formuli:
If obtained from another laboratory they should be similarly maintained for a minimum of three weeks.
% Ir postotak inhibicije prosječne specifične brzine rasta
The source of plant material and the species used for testing should always be reported.
μC srednja vrijednost μ u kontroli
The quality and uniformity of the plants used for the test will have a significant influence on the outcome of the test and should therefore be selected with care.
μT srednja vrijednost μ u tretiranoj skupini
Young, rapidly growing plants without visible lesions or discoloration (chlorosis) should be used.
Učinci na prirast određuju se na temelju mjerne varijable duljine glavnog izdanka i još triju mjernih varijabli (najbolje ukupna duljina izdanka, svježa masa, suha masa ili broj pršljena) u svakoj ispitnoj posudi na početku i na kraju ispitivanja.
Details about preparation of the test organism are given in Appendix 4.
Za svježu ili suhu masu početna biomasa određuje se na temelju uzorka ispitnih organizama uzetog iz iste šarže koja je upotrijebljena za inokulaciju ispitnih posuda.
Cultivation
Za svaku ispitnu koncentraciju i kontrolu treba izračunati srednju vrijednost prirasta i procjene varijance.
To reduce the frequency of culture maintenance (e.g. when no Myriophyllum tests are planned for a period), cultures can be held under reduced illumination and temperature (50 μE m–2 s–1, 20 ± 2 °C).
Srednji postotak inhibicije prirasta ( % Iy) za svaku tretiranu skupinu može se izračunati kako slijedi:
Details of culturing are given in Appendix 3.
% Iy postotak smanjenja prirasta
At least 14 to 21 days before testing, sufficient test organisms are transferred aseptically into fresh sterile medium and cultured for 14 to 21 days under the conditions of the test as a pre culture.
bC konačna biomasa u kontrolnoj skupini umanjena za početnu biomasu
Details for preparation of a pre-culture are given in Appendix 4.
bT konačna biomasa u tretiranoj skupini umanjena za početnu biomasu
Only one nutrient medium is recommended for Myriophyllum spicatum in a sediment-free test system, as described in Appendix 2.
Vrijeme udvostručenja
A modification of the Andrews' medium is recommended for culturing and testing with Myriophyllum spicatum as described in (1).
Da bi se odredilo vrijeme udvostručenja (Td) duljine glavnog izdanka i utvrdilo je li ispunjen taj kriterij valjanosti (vidjeti stavak 8.), dobivene podatke za kontrolne posude treba uvrstiti u sljedeću formulu:
From five separately prepared nutrient stock solutions with addition of 3 % sucrose the modified Andrews' medium will be arranged.
Td = ln 2/μ
Details about preparation of the medium are given in Appendix 2.
pri čemu je prosječna specifična brzina rasta određena kako je opisano u stavcima 50. – 52.
A tenfold concentrated, modified Andrews' medium is needed for obtaining the test solutions (by dilution as appropriate).
Prikaz krivulja koncentracija-odgovor
The composition of this medium is given in Appendix 2.
Treba izraditi krivulje koncentracija-odgovor kao prikaz odnosa srednje vrijednosti postotka inhibicije varijable odgovora (Ir ili Iy, izračunanih kako je navedeno u stavku 53.) i logaritma koncentracije ispitivane kemikalije.
Test solutions are usually prepared by dilution of a stock solution.
Procjena ECx
Stock solutions of the test chemical are normally prepared by dissolving the chemical in demineralised (i.e. distilled or deionised) water.
Procjene ECx trebale bi se temeljiti i na prosječnoj specifičnoj brzini rasta (ErCx) i na prirastu (EyCx), a svaka od njih bi se pak trebala temeljiti na duljini glavnog izdanka te eventualno drugim mjernim varijablama (najbolje ukupna duljina izdanka, svježa masa, suha masa ili broj pršljena).
The addition of the nutrients will be achieved by using the tenfold concentrated, modified Andrews' medium.
Razlog tomu je što neke kemikalije ne utječu jednako na duljinu glavnog izdanka i na druge mjerne varijable.
The stock solutions of the test chemical can be sterilised by autoclave at 121 °C for 20 minutes or by sterile filtration, provided that the sterilisation technique used does not denaturise the test chemical.
Potrebni parametri toksičnosti stoga su četiri vrijednosti ECx za svaku izračunanu razinu inhibicije x: ErCx (duljina glavnog izdanka), ErCx (najbolje ukupna duljina izdanka, svježa masa, suha masa ili broj pršljena), EyCx (duljina glavnog izdanka) i EyCx (najbolje ukupna duljina izdanka, svježa masa, suha masa ili broj pršljena).
Test solutions can also be prepared in sterile demineralised water or medium, under sterile conditions.
Treba napomenuti da vrijednosti ECx izračunane primjenom tih dviju varijabli odgovora nisu usporedive i tu razliku treba uzeti u obzir pri korištenju rezultatima ispitivanja.
The thermo-stability and the adsorption on different surfaces should the taken into account in the selection of the sterilisation procedure of the stock solutions of the test chemical.
Uz poštovanje ispitnih uvjeta ove ispitne metode, vrijednosti ECx na temelju prosječne specifične brzine rasta (ErCx) u većini će slučajeva biti više od rezultata na temelju prirasta (EyCx) zbog razlike u matematičkoj osnovi tih dvaju pristupa.
Because of that, it is recommended that the stock solutions be prepared under sterile conditions, i.e. using sterile material for dissolving the test chemical under sterile conditions (e.g. flame sterilisation, laminar-flow hoods, etc.) into sterile water.
Tu razliku ne treba tumačiti kao razliku u osjetljivosti tih dviju varijabli odgovora; jednostavno je riječ o matematički različitim vrijednostima.
This technique of preparation of sterile stock solutions is valid for both substances and mixtures.
Statistički postupci
The highest tested concentration of the test chemical should normally not exceed its water solubility under the test conditions.
Cilj je regresijskom analizom dobiti kvantitativan odnos koncentracija-odgovor.
For test chemicals of low water solubility it may be necessary to prepare a concentrated stock solution or dispersion of the chemical using an organic solvent or dispersant in order to facilitate the addition of accurate quantities of the test chemical to the test medium and aid in its dispersion and dissolution.
Ako se provede linearizacijska transformacija podataka odgovora, npr. s pomoću probit, logit ili Weibullova modela (7.), može se primijeniti ponderirana linearna regresija; ipak, prednost se daje postupcima nelinearne regresije kojima se bolje rješavaju neizbježne nepravilnosti podataka i odstupanja od pravilnih distribucija.
Every effort should be made to avoid the use of such materials.
S približavanjem izostanku inhibicije ili potpunoj inhibiciji te se nepravilnosti transformacijom mogu i dodatno povećati te tako otežati analizu (7.).
There should be no phytotoxicity resulting from the use of auxiliary solvents or dispersants.
Treba napomenuti da su standardne metode analize za vrijednosti probit, logit ili Weibull namijenjene kvantalnim podacima (npr. smrtnost ili preživljavanje) i moraju se posebno prilagoditi da bi se mogle primijeniti na podatke o brzini rasta i prirastu.
For example, commonly used solvents which do not cause phytotoxicity at concentrations up to 100 μl/l, include acetone and dimethylformamide.
Konkretni postupci za određivanje vrijednosti ECx iz kontinuiranih podataka mogu se pronaći u literaturi (8., 9. i 10.).
If a solvent or dispersant is used, its final concentration should be reported and kept to a minimum (≤ 100 μl/l), and all treatments and controls should contain the same concentration of solvent or dispersant.
Za svaku varijablu odgovora koja se analizira treba izračunati procjene vrijednosti ECx na temelju odnosa koncentracija-odgovor.
Further guidance on the use of dispersants is given in (5).
Po mogućnosti, za sve procjene treba odrediti granice pouzdanosti od 95 %.
Test and control groups
Valjanost podudaranja podataka odgovora s regresijskim modelom trebalo bi procijeniti grafički ili statistički.
Prior knowledge of the toxicity of the test chemical to Myriophyllum spicatum from a range-finding test will help in selecting suitable test concentrations.
Regresijsku analizu trebalo bi provesti na temelju odgovora pojedinačnih ponovljenih uzoraka, a ne na temelju srednjih vrijednosti tretiranih skupina.
In the definitive toxicity test, there should normally be five (like in the Lemna growth inhibition test, Chapter C.26 of this Annex) to seven test concentrations arranged in a geometric series; they should be chosen in order that the NOEC and EC50 values are bracketed by the concentration range (see below).
Ako raspoloživi regresijski modeli / regresijske metode nisu prikladni/-e za te podatke, procjene EC50 i granice pouzdanosti mogu se dobiti i linearnom interpolacijom sa samonadopunjavanjem (bootstrapping) (10.).
Preferably the separation factor between test concentrations should not exceed 3,2; however, a larger value may be used where the concentration-response curve is flat.
Za procjenu LOEC-a, a time i NOEC-a, potrebno je usporediti srednje vrijednosti tretiranih skupina primjenom tehnika analize varijance (ANOVA).
Justification should be provided when fewer than five concentrations are used.
Zatim srednju vrijednost za svaku koncentraciju treba usporediti sa srednjom vrijednošću kontrole primjenom odgovarajuće metode višestruke usporedbe ili testa trenda.
At least five replicates should be used at each test concentration.
Može biti koristan Dunnettov ili Williamsov test (12., 13., 14., 15., 16.).
In setting the range of test concentrations (for range-finding and/or for the definitive toxicity test), the following should be considered:
Treba provjeriti vrijedi li pretpostavka homogenosti varijanci ANOVA-e.
To determine an ECx, test concentrations should bracket the ECx value to ensure an appropriate level of confidence.
To se može procijeniti grafički ili formalnim testom (15.).
For example, if estimating the EC50, the highest test concentration should be greater than the EC50 value.
Prikladni su Leveneov ili Bartlettov test.
If the EC50 value lies outside of the range of test concentrations, associated confidence intervals will be large and a proper assessment of the statistical fit of the model may not be possible.
Ako pretpostavka homogenosti varijanci nije ispunjena, to se ponekad može ispraviti logaritamskom transformacijom podataka.
If the aim is to estimate the LOEC/NOEC, the lowest test concentration should be low enough so that growth is not significantly less than that of the control.
Ako je heterogenost varijanci prevelika i ne može se ispraviti transformacijom, treba razmotriti mogućnost analize metodama kao što su Jonckheereovi ‚step-down’ testovi trenda.
In addition, the highest test concentration should be high enough so that growth is significantly lower than that in the control.
Dodatne smjernice za određivanje NOEC-a mogu se pronaći u literaturi (10.).
If this is not the case, the test will have to be repeated using a different concentration range (unless the highest concentration is at the limit of solubility or the maximum required limit concentration, e.g. 100 mg/l).
Novije znanstvene spoznaje rezultirale su preporukom da se pojam NOEC-a napusti i zamijeni procjenama ECx dobivenima regresijom.
Every test should include controls consisting of the same nutrient medium, test organism (choosing plant material as homogeneous as possible, fresh lateral branches from pre-cultures, shortened to 2,5 cm from base), environmental conditions and procedures as the test vessels but without the test chemical.
Za ovo ispitivanje s klasastim krocnjem nije utvrđena primjerena vrijednost x.
If an auxiliary solvent or dispersant is used, an additional control treatment with the solvent/dispersant present at the same concentration as that in the vessels with the test chemical should be included.
Ipak, čini se da je primjeren raspon od 10 do 20 % (ovisno o odabranoj varijabli odgovora), a poželjno je da se navedu obje vrijednosti, EC10 i EC20, te njihove granice pouzdanosti.
The number of replicate control vessels (and solvent vessels, if applicable) should be at least ten.
Izvješćivanje
If determination of NOEC is not required, the test design may be altered to increase the number of concentrations and reduce the number of replicates per concentration.
tvar s više sastojaka, tvar nepoznatog ili promjenjivog sastava, složeni reakcijski proizvod ili biološki materijal (UVCB tvar) ili smjesa:
However, in any case the number of control replicates should be at least ten.
Ispitne vrste
Exposure
znanstveni naziv i podrijetlo.
Fresh lateral branches from pre-culture shortened to 2,5 cm from base are assigned randomly to the test vessels under aseptic conditions; each test vessel should contain one 2,5 cm lateral branch that should have an apical meristem on one end.
ispitni postupak (statički ili polustatički),
The chosen plant material should be the same quality in each test vessel.
datum početka i trajanje ispitivanja,
A randomised design for location of the test vessels in the incubator is required to minimise the influence of spatial differences in light intensity or temperature.
ispitni medij,
A blocked design or random repositioning of the vessels (or repositioning more frequently) when observations are made is also required.
opis plana ispitivanja: ispitne posude i pokrovi, količine otopina, duljina glavnog izdanka po ispitnoj posudi na početku ispitivanja,
If a preliminary stability test shows that the test chemical concentration cannot be maintained (i.e. the measured concentration falls below 80 % of the measured initial concentration) over the test duration (14 days), a semi-static test regime is recommended.
ispitne koncentracije (nazivne odnosno izmjerene, prema potrebi) i broj ponovljenih uzoraka po koncentraciji,
In this case, the plants should be exposed to freshly prepared test and control solutions on at least one occasion during the test (e.g. day 7).
način pripreme matičnih i ispitnih otopina, uključujući eventualnu upotrebu otapala ili dispergenata,
The frequency of exposure to fresh medium will depend on the stability of the test chemical; a higher frequency may be needed to maintain near-constant concentrations of highly unstable or volatile chemicals.
temperatura tijekom ispitivanja,
The exposure scenario through a foliar application (spray) is not covered in this test method.
izvor svjetlosti, intenzitet i homogenost svjetlosti,
Warm and/or cool white fluorescent lighting should be used to provide light irradiance in the range of about of 100-150 μE m–2 s–1 when measured as a photosynthetically active radiation (400-700 nm) at points the same distance from the light source as the bottom of the test vessels (equivalent ca. 6000 to 9000 lux) and using a light-dark cycle of 16:8 h.
pH-vrijednosti ispitnih i kontrolnih medija,
Spherical sensors (which respond to light from all angles above and below the plane of measurement) and “cosine” sensors (which respond to light from all angles above the plane of measurement) are preferred to unidirectional sensors, and will give higher readings for a multi-point light source of the type described here.
metode određivanja duljine glavnog izdanka i drugih mjernih varijabli, npr. ukupne duljine bočnih grana, ukupne duljine izdanka, ukupne duljine korijena, svježe mase, suhe mase ili broja pršljena,
The temperature in the test vessels should be 23 ± 2 °C.
stanje kulture (sterilna ili nesterilna) u svakoj ispitnoj i kontrolnoj posudi pri svakom promatranju,
See (5) for further guidance.
neobrađeni podaci: duljina glavnog izdanka i druge mjerne varijable u svakoj ispitnoj i kontrolnoj posudi pri svakom promatranju i svakoj analizi,
The test is terminated 14 days after the plants are transferred into the test vessels.
srednje vrijednosti i standardne devijacije za svaku mjernu varijablu,
Measurements and analytical determinations
krivulje rasta za svaku mjernu varijablu,
At the start of the test, the main shoot length of test organism is 2,5 cm (see paragraph 29); it is measured with a ruler (see Appendix 4) or by photography and image analysis.
izračunane varijable odgovora za svaki tretirani ponovljeni uzorak, uključujući srednje vrijednosti i koeficijente varijacije za ponovljene uzorke,
The main shoot length of test organism appearing normal or abnormal needs to be determined at the beginning of the test, at least once during the 14-day exposure period and at test termination.
grafički prikaz odnosa koncentracija-učinak,
Note: As an alternative for those who do not have image analysis, if the workbench is sterilised prior to addition of plants to test vessels, a sterile ruler can also be used to measure the length of the main shoot at test initiation and during the test.
procjene krajnjih točaka toksičnosti za varijable odgovora, npr.
Changes in plant development, e.g. in deformation in the shoots, appearance, indication of necrosis, chlorosis, break-up or loss of buoyancy and in root length and appearance, should be noted.
EC50, EC10, EC20, i pripadajući intervali pouzdanosti; ako se izračunavaju LOEC i NOEC, njihove vrijednosti i statističke metode koje su upotrijebljene za izračun,
Significant features of the test medium (e.g. presence of undissolved material, growth of algae, fungi and bacteria in the test vessel) should also be noted.
ako se primjenjuje ANOVA, jačina učinka koji se može utvrditi (npr. najmanja značajna razlika),
In addition to determinations of main shoot length during the test, effects of the test chemical on three (or more) of the following measurement variables should be also assessed:
svaka stimulacija rasta u bilo kojoj tretiranoj skupini,
Total lateral branches length
svi vidljivi znakovi fitotoksičnosti i zapažanja u ispitnim otopinama,
Total shoot length
rasprava o rezultatima, uključujući i sve utjecaje na rezultate ispitivanja nastale uslijed odstupanja od ove ispitne metode.
Total root length
ASTM Designation E 1913-04, Standard Guide for Conducting Static, Axenic, 14-Day Phytotoxicity Tests in Test Tubes with the Submersed Aquatic Macrophyte, Myriophyllum sibiricum Komarov.
Fresh weight
Maletzki, D. et al.
Dry weight
(2010), Myriophyllum spicatum als ökotoxikologischer Testorganismus: Methodenentwicklung eines sedimentfreien Testsystems und erste Ergebnisse mit 3,5-Dichlorphenol, Umweltwiss Schadst Forsch, No.
Number of whorls
22, str.
Note 1:
702-710.
The observations made during the range-finding test could help in selecting relevant additional measurements among the six variables listed above.
Poglavlje C.26. ovog Priloga: Test inhibicije rasta Lemna sp.
Note 2:
OECD (2014), ‚Myriophyllum spicatum Toxicity Test: Results of an inter-laboratory ring test using a sediment-free test system’, OECD Environment, Health and Safety Publications (EHS), Series Testing and Assessment, No. 205, OECD Publishing, Paris.
The determination of the fresh and dry weights (parameters iv and v) is highly desirable.
OECD (2000), ‚Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures’, OECD Environment, Health and Safety Publications (EHS), Series on Testing and Assessment, No. 23, OECD Publishing, Paris.
Note 3:
Poglavlje C.51. ovog Priloga: Ispitivanje toksičnosti na klasastom krocnju (Myriophyllum spicatum) u sustavu voda-sediment.
Due to the fact that sucrose and light (exposure of roots to light during the test) may have an influence on auxin (plant growth hormone) transport carriers, and that some chemicals may have an auxin-type mode of action, the inclusion of root endpoints (parameter iii) is questionable.
Christensen, E.R., N. Nyholm (1984), Ecotoxicological Assays with Algae: Weibull Dose-Response Curves, Environmental Science & Technology, Vol.
Note 4:
18/9, 713-718.
The ring test results show high coefficients of variation (> 60 %) for the total lateral branch length (parameter i).
Bruce, R.D., D.J.
Total lateral branch length is in any case encompassed within the total shoot length measurement (parameter ii) which shows more acceptable coefficients of variation of < 30 %.
OECD (2006), ‚Current Approaches in the Statistical Analysis of Ecotoxicity Data: A Guidance to Application’, OECD Environment, Health and Safety Publications (EHS), Series on Testing and Assessment, No. 54, OECD Publishing, Paris.
Note 5:
Dunnett, C.W.
Resulting from the above considerations, the recommended main measurement endpoints are: total shoot length, fresh weight and dry weight (parameters ii, iv and v); parameter vi — number of whorls — is left to the experimenter's judgment.
, A multiple comparisons procedure for comparing several treatments with a control, Journal of the American Statistical Association, Vol.
Main shoot length and number of whorls have an advantage, in that they can be determined for each test and control vessel at the start, during, and at the end of the test by photography and image analysis, although a (sterile) ruler can also be used.
50/272, str. 1096-1121.
Total lateral branches length, total root length (as a sum of all lateral branches or roots) and total shoot length (as a sum of main shoot length and total lateral branches length) can be measured with a ruler at the end of exposure.
, New tables for multiple comparisons with a control, Biometrics, Vol.