natgillin/translations-raw huggingface dataset:
Til beregning af et estimat for LOEC og dermed NOEC må middelværdierne for behandlingerne sammenholdes ved hjælp af variansanalyse.
Continuous warm or cool white fluorescent lighting should be used to provide a light intensity selected from the range of 85-135 μE · m– 2s– 1 when measured in a photosynthetically active radiation (400-700 nm) at points the same distance from the light source as the Lemna fronds (equivalent to 6500-10000 lux).
Dunnetts eller Williams' test kan være nyttig (18)(19)(20)(21).
Any differences from the selected light intensity over the test area should not exceed the range of ± 15 %.
Denne vurdering kan foretages grafisk eller ved en formel test (22).
The method of light detection and measurement, in particular the type of sensor, will affect the measured value.
Supplerende vejledning for bestemmelse af NOEC er givet i (16).
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.
Der er ikke fastlagt en passende værdi af x til denne Lemna-test.
The temperature in the test vessels should be 24 ± 2 °C.
Et område på 10 til 20 % synes imidlertid at være passende (afhængigt af den valgte responsvariabel), og det må foretrækkes at rapportere både EC10 og EC20.
However, deviation of more than 1,5 units would not invalidate the test when it can be shown that validity criteria are met.
Rapportering
Additional care is needed on pH drift in special cases such as when testing unstable chemicals or metals.
fysisk tilstand og fysisk-kemiske egenskaber, herunder vandopløselighed
See (8) for further guidance.
videnskabeligt navn, klon (hvis den kendes) og kilde.
Duration
den anvendte testmetode (statisk, semistatisk eller gennemstrømningstest)
The test is terminated 7 days after the plants are transferred into the test vessels.
testsubstrat
At the start of the test, frond number in the test vessels is counted and recorded, taking care to ensure that protruding, distinctly visible fronds are accounted for.
beskrivelse af forsøgsdesign:
Frond numbers appearing normal or abnormal, need to be determined at the beginning of the test, at least once every 3 days during the exposure period (i.e. on at least 2 occasions during the 7 day period), and at test termination.
testbeholdere og låg, opløsningsvolumener, antal kolonier og blade pr. testbeholder ved testens start
Changes in plant development, e.g. in frond size, appearance, indication of necrosis, chlorosis or gibbosity, colony break-up or loss of buoyancy, and in root length and appearance, should be noted.
testkoncentrationer (nominelle eller målte) og antal replikater for hver koncentration
Significant features of the test medium (e.g. presence of undissolved material, growth of algae in the test vessel) should also be noted.
metoder til fremstilling af stam- og testopløsninger, herunder eventuelle opløsnings- og dispergeringsmidler
In addition to determinations of frond number during the test, effects of the test chemical on one (or more) of the following measurement variables are also assessed:
testtemperatur
total frond area,
lyskilde, lysintensitet og -homogenitet
dry weight,
test- og kontrolsubstraternes pH-værdi
fresh weight.
testkemikaliekoncentrationer og analysemetoder med fyldestgørende oplysninger om kvalitetsvurdering (valideringsundersøgelser, standardafvigelser eller konfidensgrænser for analyserne)
Total frond area has an advantage, in that it can be determined for each test and control vessel at the start, during, and at the end of the test.
metoder til bestemmelse af bladantal og andre målevariable, f.eks. tørvægt, frisk vægt og bladareal
Dry or fresh weight should be determined at the start of the test from a sample of the inoculum culture representative of what is used to begin the test, and at the end of the test with the plant material from each test and control vessel.
alle fravigelser af nærværende testmetode.
If frond area is not measured, dry weight is preferred over fresh weight.
rådata:
Total frond area, dry weight and fresh weight may be determined as follows:
antal blade og andre målevariable i hver test- og kontrolbeholder ved hver observation og hver analyse
Total frond area:
gennemsnit og standardafvigelse for hver målevariabel
The total frond area of all colonies may be determined by image analysis.
vækstkurver for hver koncentration (brug af logaritmisk transformerede målevariable anbefales, se punkt 55)
A silhouette of the test vessel and plants can be captured using a video camera (i.e. by placing the vessel on a light box) and the resulting image digitised.
fordoblingstid/væksthastighed i kontrollen, baseret på bladantal
By calibration with flat shapes of known area, the total frond area in a test vessel may then be determined.
grafisk fremstilling af sammenhængen mellem koncentration og virkning
Care should be taken to exclude interference caused by the rim of the test vessel.
estimater for toksicitetsendepunkter for responsvariablene, f.eks. EC50, EC10, EC20, med tilhørende konfidensintervaller.
An alternative but more laborious approach is to photocopy test vessels and plants, cut out the resulting silhouette of colonies and determine their area using a leaf area analyser or graph paper.
LOEC og/eller NOEC (hvis disse er beregnet) og de statistiske metoder, der anvendt til at beregne dem
Other techniques (e.g. paper weight ratio between silhouette area of colonies and unit area) may also be appropriate.
ethvert visuelt tegn på fytotoksicitet samt observationer af testopløsningerne
Dry weight:
diskussion af resultaterne, herunder den eventuelle betydning af fravigelser fra denne testmetode.
All colonies are collected from each of the test vessels and rinsed with distilled or deionised water.
Test for Measuring the Inhibition of Growth Using the Freshwater Macrophyte, Lemna minor, EPS 1/RM/37 — s. 120 ff.
They are blotted to remove excess water and then dried at 60 °C to a constant weight.
Non-Target Plant Testing and Evaluation, Canadian Wildlife Service, Technical Report Series No. 145.
Any root fragments should be included.
Følgende definitioner og forkortelser anvendes i denne testmetode:
The dry weight should be expressed to an accuracy of at least 0,1 mg.
Biomasse er tørvægten af det levende materiale, som er til stede i en population.
Fresh weight:
I denne test måles typisk surrogater for biomasse, f.eks. bladantal eller bladareal, og betegnelsen »biomasse« benyttes således også for sådanne surrogatmålinger.
All colonies are transferred to pre-weighed polystyrene (or other inert material) tubes with small (1 mm) holes in the rounded bottoms.
Chlorose er gulning af bladvæv.
The tubes are then centrifuged at 3000 rpm for 10 minutes at room temperature.
Klon er en organisme eller celle, der er opstået af en enkelt celle ved vegetativ formering.
Tubes, containing the now dried colonies, are re-weighed and the fresh weight is calculated by subtracting the weight of the empty tube.
Individer fra samme klon er derfor genetisk identiske.
Frequency of measurements and analytical determinations
Koloni er en samling af moder- og datterblade (sædvanligvis 2 til 4), der sidder fast på hinanden.
If a static test design is used, the pH of each treatment should be measured at the beginning and at the end of the test.
Betegnes undertiden en plante.
If a semi-static test design is used, the pH should be measured in each batch of “fresh” test solution prior to each renewal and also in the corresponding “spent” solutions.
ECx er den koncentration af testkemikalie opløst i testsubstratet, der medfører en væksthæmning på x % (f.eks. 50 %) af Lemna i løbet af en nærmere bestemt eksponeringsperiode (som udtrykkeligt skal angives, hvis den er forskellig fra den fulde eller normale testperiode).
Light intensity should be measured in the growth chamber, incubator or room at points the same distance from the light source as the Lemna fronds.
For klart at skelne mellem EC-værdier afledt af henholdsvis væksthastighed og udbytte anvendes betegnelsen »ErC« for væksthastighed (rate) og »EyC« for udbytte (yield), efterfulgt af den anvendte målevariabel, f.eks. »ErC (bladantal)«.
Measurements should be made at least once during the test.
Gennemstrømningstest er en test, hvor testopløsningen udskiftes kontinuerligt.
The temperature of the medium in a surrogate vessel held under the same conditions in the growth chamber, incubator or room should be recorded at least daily.
Blad er en individuel/enkelt »bladlignende« struktur hos andemadplanten.
During the test, the concentrations of the test chemical are determined at appropriate intervals.
Det er den mindste individuelle enhed, der kan formere sig.
In static tests, the minimum requirement is to determine the concentrations at the beginning and at the end of the test.
Opsvulmethed er blade, der fremtræder udbulede eller opsvulmede.
In semi-static tests where the concentration of the test chemical is not expected to remain within ± 20 % of the nominal concentration, it is necessary to analyse all freshly prepared test solutions and the same solutions at each renewal (see paragraph 33).
Vækst er en stigning i en målevariabel i løbet af testperioden, f.eks. bladantal, tørvægt, vådvægt eller bladareal.
However, for those tests where the measured initial concentration of the test chemical is not within ± 20 % of nominal but where sufficient evidence can be provided to show that the initial concentrations are repeatable and stable (i.e. within the range 80 - 120 % of the initial concentration), chemical determinations may be carried out on only the highest and lowest test concentrations.
Målevariable er enhver type variabel, der måles for at udtrykke endepunktet for en test ved hjælp af en eller flere forskellige responsvariable.
In all cases, determination of test chemical concentrations prior to renewal need only be performed on one replicate vessel at each test concentration (or the contents of the vessels pooled by replicate).
I denne metode er bladantal, bladareal, frisk vægt og tørvægt målevariable.
If a flow-through test is used, a similar sampling regime to that described for semi-static tests, including analysis at the start, mid-way through and at the end of the test, is appropriate, but measurement of “spent” solutions is not appropriate in this case.
Monokultur er en kultur med en planteart.
In this type of test, the flow-rate of diluent and test chemical or test chemical stock solution should be checked daily.
Nekrose er dødt (dvs. hvidt eller gennemblødt) bladvæv.
If the deviation from the nominal or measured initial concentration is not within ± 20 %, analysis of the results should be based on the geometric mean concentration during exposure or models describing the decline of the concentration of the test chemical (8).
Fænotype er de observerbare kendetegn for en organisme, som er bestemt af samspillet mellem dens gener og dens omgivelser.
All previously described test conditions and validity criteria apply to a limit test, with the exception that the number of treatment replicates should be doubled.
Responsvariable er variable, som anvendes til vurdering af den toksiske virkning og er afledt af målte parametre, der beskriver biomassen ved forskellige beregningsmetoder.
Growth in the control and treatment group may be analysed using a statistical test to compare means, e.g. a Student's t-test.
I nærværende testmetode er væksthastighed og udbytte responsvariable, der er afledt af målevariable som bladantal, bladareal, frisk vægt eller tørvægt.
Doubling time
Semistatisk test (udskiftningstest) er en test, hvor testopløsningen udskiftes med bestemte intervaller i løbet af testen.
To determine the doubling time (Td) of frond number and adherence to this validity criterion by the study (paragraph 12), the following formula is used with data obtained from the control vessels:
Statisk test er en testmetode, hvor testopløsningen ikke udskiftes under testen.
where μ is the average specific growth rate determined as described in paragraphs 54-55.
Testkemikalie stoffer eller blandinger, der testes med denne testmetode.
The purpose of the test is to determine the effects of the test chemical on the vegetative growth of Lemna.
Endepunkt for en test er den generelle faktor, som testkemikaliet fremkalder en ændring af i forhold til kontrollen som mål for testen.
This Test Method describes two response variables, as different jusidictions have different preferences and regulatory needs.
I denne testmetode er endepunktet væksthæmning, som kan udtrykkes ved forskellige responsvariable, der er baseret på en eller flere målevariable.
Average specific growth rate:
Testsubstrat er det komplette, syntetiske næringssubstrat, hvorpå testplanterne vokser, når de eksponeres for testkemikaliet.
this response variable is calculated on the basis of changes in the logarithms of frond numbers, and in addition, on the basis of changes in the logarithms of another measurement parameter (total frond area, dry weight or fresh weight) over time (expressed per day) in the controls and each treatment group.
Udbytte er værdien af en målevariabel for biomasse ved eksponeringsperiodens slutning minus værdien af samme variabel ved eksponeringsperiodens start.
It is sometimes referred to as relative growth rate (12).
Beskrivelse af Lemna spp.
Yield:
Vandplanten med trivialnavnet andemad, Lemna spp., tilhører familien Lemnaceae, der omfatter en række arter, som er udbredt over hele verden og består af fire slægter.
this response variable is calculated on the basis of changes in frond number, and in addition, on the basis of changes in another measurement parameter (total frond area, dry weight or fresh weight) in the controls and in each treatment group until the end of the test.
Deres respektive udseende og taksonomi er beskrevet udtømmende (1)(2).
ECx values based upon average specific growth rate (ErCx) will generally be higher than results based upon yield (EyCx) if the test conditions of this Test Method are adhered to, due to the mathematical basis of the respective approaches.
Lemna gibba og L. minor er repræsentative arter fra tempererede områder og finder almindelig anvendelse i toksicitetstest.
The concept of average specific growth rate is based on the general exponential growth pattern of duckweed in non-limited cultures, where toxicity is estimated on the basis of the effects on the growth rate, without being dependent on the absolute level of the specific growth rate of the control, slope of the concentration-response curve or on test duration.
Begge arter har flydende eller nedsænket skiveformet stilk (blad) og en meget tynd rod, der udgår fra midten af undersiden på hvert blad.
EyCx is dependent on the specific growth rate of the duckweed species used in each test and on the maximum specific growth rate that can vary between species and even different clones.
Lemna spp. danner sjældent blomster, og planterne formerer sig vegetativt ved at skyde nye blade (3).
This response variable should not be used for comparing the sensitivity to toxicants among duckweed species or even different clones.
De unge planter er blegere og har kortere rødder end de ældre planter og består af to-tre blade af forskellig størrelse.
While the use of average specific growth rate for estimating toxicity is scientifically preferred, toxicity estimates based on yield are also included in this Test Method to satisfy current regulatory requirements in some jurisdictions.
Lemnas enkle struktur, ukønnede formering og korte generationstid gør planten meget velegnet til laboratorietest (4)(5).
Toxicity estimates should be based on frond number and one additional measurement variable (total frond area, dry weight or fresh weight), because some chemicals may affect other measurement variables much more than the frond number.
På grund af artsbetingede forskelle i følsomhed er sammenligning af følsomhed kun gyldig inden for samme art.
This effect would not be detected by calculating frond number only.
Opbevaring af stamkulturen
The number of fronds as well as any other recorded measurement variable, i.e. total frond area, dry weight or fresh weight, are tabulated together with the concentrations of the test chemical for each measurement occasion.
Ved lavere temperatur (4-10 °C) kan stamkulturer holdes i længere tid uden at skulle gendannes.
Subsequent data analysis e.g. to estimate a LOEC, NOEC or ECx should be based on the values for the individual replicates and not calculated means for each treatment group.
Næringssubstratet til Lemna kan være det samme som det, der anvendes ved testning, men andre næringsrige substrater kan anvendes til stamkulturer.
The average specific growth rate for a specific period is calculated as the logarithmic increase in the growth variables -frond numbers and one other measurement variable (total frond area, dry weight or fresh weight) — using the formula below for each replicate of control and treatments:
Med jævne mellemrum udtages et antal unge, lysegrønne planter og overføres aseptisk til nye dyrkningsbeholdere med frisk substrat.
average specific growth rate from time i to j
Under de her foreslåede køligere forhold kan dyrkning af subkulturer ske med op til tre måneders mellemrum.
measurement variable in the test or control vessel at time i
Der skal anvendes kemisk rene (syrevaskede) og sterile dyrkningsbeholdere af glas samt aseptisk håndtering.
measurement variable in the test or control vessel at time j
Hvis en stamkultur bliver kontamineret med f.eks. alger eller svampe, må der foretages indgreb til at eliminere kontaminanten.
t time period from i to j
For algers og de fleste andre kontaminanters vedkommende kan dette opnås ved overfladesterilisation.
The average specific growth rate should be calculated for the entire test period (time “i” in the above formula is the beginning of the test and time “j” is the end of the test).
Substrater
mean value for μ in the control
Der anbefales forskellige næringssubstrater til L. minor og L. gibba.
mean value for μ in the treatment group
Til L. minor anbefales et modificeret substrat efter Swedish Standard (SIS), mens substratet 20X AAP anbefales til L. gibba.
Effects on yield are determined on the basis of two measurement variables, frond number and one other measurement variable (total frond area, dry weight or fresh weight) present in each test vessel at the start and at the end of the test.
Nedenfor er sammensætningen af de to substrater angivet.
For dry weight or fresh weight, the starting biomass is determined on the basis of a sample of fronds taken from the same batch used to inoculate the test vessels (see paragraph 20).
Lemna-næringssubstrat efter Swedish Standard (SIS)
The mean percent inhibition in yield (% Iy) may be calculated for each treatment group as follows:
Stamopløsning I-V steriliseres ved autoklavering (120 °C, 15 min.) eller ved membranfiltrering (porediameter ca. 0,2 μm).
percent reduction in yield
Sterile stamopløsninger skal opbevares køligt og mørkt.
final biomass minus starting biomass for the control group
Stamopløsning I-V skal kasseres efter seks måneder, mens holdbarheden for stamopløsning VI (og valgfrit VII) er en måned.
final biomass minus starting biomass in the treatment group
Stamopløsning nr.
Plotting concentration-response curves
Stof
Concentration-response curves relating mean percentage inhibition of the response variable (Ir, or Iy calculated as shown in paragraph 56 or 57) and the log concentration of the test chemical should be plotted.
Koncentration i færdigt substrat
ECx estimation
Færdigt substrat
Estimates of the ECx (e.g., EC50) should be based upon both average specific growth rate (ErCx) and yield (EyCx), each of which should in turn be based upon frond number and one additional measurement variable (total frond area, dry weight, or fresh weight).
Koncentration
This is because there are test chemicals that impact frond number and other measurement variables differently.
ZnSO4 · 7H2O
The desired toxicity parameters are therefore four ECx values for each inhibition level x calculated:
CuSO4 · 5H2O
ErCx (frond number);
Co(NO3)2 · 6H2O
ErCx (total frond area, dry weight, or fresh weight);
Til fremstilling af en liter SIS-substrat tilsættes følgende til 900 ml deioniseret vand:
EyCx (frond number);
10 ml stamopløsning I
and EyCx (total frond area, dry weight, or fresh weight).
5 ml stamopløsning II
It is possible to use a weighted linear regression after having performed a linearising transformation of the response data, for instance into probit or logit or Weibull units (13), but non-linear regression procedures are preferred techniques that better handle unavoidable data irregularities and deviations from smooth distributions.
5 ml stamopløsning III
Approaching either zero or total inhibition such irregularities may be magnified by the transformation, interfering with the analysis (13).
5 ml stamopløsning IV
It should be noted that standard methods of analysis using probit, logit, or Weibull transforms are intended for use on quantal (e.g. mortality or survival) data, and must be modified to accommodate growth rate or yield data.
1 ml stamopløsning V
Specific procedures for determination of ECx values from continuous data can be found in (14), (15), and (16).
5 ml stamopløsning VI
estimates and confidence limits may also be obtained using linear interpolation with bootstrapping (17), if available regression models/methods are unsuitable for the data.
1 ml stamopløsning VII (valgfrit)
For estimation of the LOEC and hence the NOEC, it is necessary to compare treatment means using analysis of variance (ANOVA) techniques.
Bemærk:
Dunnett's or Williams'test may be useful (18)(19)(20)(21).
Til visse testkemikalier kan der yderligere være brug for en stamopløsning VII (MOPS-buffer) (se punkt 11).
This assessment may be performed graphically or by a formal test (22).
Der fremstilles fem næringssubstrat-stamopløsninger (A1, A2, A3, B og C) til 20X-AAP-substrat, idet der anvendes kemikalier af reagenskvalitet.
Additional guidance on determining the NOEC can be found in (16).
Substratet filtreres gennem et membranfilter med en porediameter på ca. 0,2 μm over i en steril beholder.
An appropriate value for x has not been established for this Lemna test.
Det næringssubstrat, der skal bruges til testningen, skal fremstilles 1-2 dage før brug, for at pH kan stabiliseres.
However, a range of 10 to 20 % appears to be appropriate (depending on the response variable chosen), and preferably both the EC10 and EC20 should be reported.
Næringssubstratets pH skal kontrolleres før brug og om nødvendigt justeres igen med 0,1 M eller 1 M NaOH eller HCl som beskrevet ovenfor.
Reporting
Den teoretisk mest passende endelige bicarbonatkoncentration (som vil overflødiggøre nævneværdig pH-justering) er 15 mg/l, ikke 300 mg/l.
physical nature and physical-chemical properties, including water solubility limit;
Den traditionelle brug af 20X-AAP-substrat og ringtesten i forbindelse med denne vejledning er imidlertid baseret på 300 mg/l.
scientific name, clone (if known) and source.
Stof
test procedure used (static, semi-static or flow-through);
EDTA · 2H2O
test medium;
STEINBERG-substrat (efter ISO 20079)
description of the experimental design:
Koncentrationer og stamopløsninger
test vessels and covers, solution volumes, number of colonies and fronds per test vessel at the beginning of the test;
Næringssubstratet fremstilles af stamopløsninger eller af det 10-dobbelt koncentrerede substrat, som har den maksimale koncentration af substrat uden udfældning.
temperature during the test;
pH-stabiliseret STEINBERG-substrat (modificeret efter Altenburger)
light source, light intensity and homogeneity;
Næringssubstrat
pH values of the test and control media;
Makronæringsstoffer
test chemical concentrations and the method of analysis with appropriate quality assessment data (validation studies, standard deviations or confidence limits of analyses);
Molvægt
methods for determination of frond number and other measurement variables, e.g. dry weight, fresh weight or frond area;
Mikronæringsstoffer
all deviations from this Test Method.
EDTA-dinatrium-dihydrat
raw data: