natgillin/translations-raw huggingface dataset:
Bunden af røret dækkes med et stykke filterpapir, der påfyldes vand, og røret placeres herefter på et stativ i et vandbad.
The worms can be cultured in large aquaria (57 - 80 l) at 23 °C with a 16 L:8 D photoperiod (100 – 1000 lx) using daily renewed natural water (45 - 50 l per aquarium).
Dyrkning af Hypoaspis (Geolaelaps) aculeifer, fodermider og synkronisering af kultur
The substrate is prepared by cutting unbleached brown paper towels into strips, which may then be blended with culture water for a few seconds to result in small pieces of paper substrate.
Dyrkning af Hypoaspis (Geolaelaps) aculeifer:
This substrate can then directly be used in the Lumbriculus culture aquaria by covering the bottom area of the tank, or be stored frozen in deionised water for later use.
Kulturer kan opbevares i plastbeholdere eller glas fyldt med en blanding af gips og kulpulver (i forholdet 9:1).
New substrate in the tank will generally last for approximately two months.
Gipsen kan holdes fugtig ved at tilsætte et par dråber destilleret eller deioniseret vand, hvis det er nødvendigt.
Each worm culture is started with 500 – 1000 worms, and fed a 10 ml suspension containing 6 g of trout starter food 3 times per week under renewal or flow-through conditions.
De optimale dyrkningstemperaturer er på 20 °C ± 2 °C;
Static or semi-static cultures should receive lower feeding rates to prevent bacterial and fungal growth. .
lys/mørke-forhold er ikke relevante for denne art. Byttedyr kan være Typrophagus putrescentiae- eller Caloglyphus sp.-mider (fodermider skal håndteres forsigtigt, da de kan give allergi hos mennesker), men nematoder, enchytraeider og collemboler er også egnede som byttedyr.
Under these conditions the number of individuals in the culture generally doubles in approximately 10 to 14 d.
Deres kilde skal registreres.
Alternatively Lumbriculus variegatus can also be cultured in a system consisting of a layer of quartz sand as used for the artificial sediment (1 - 2 cm depth), and reconstituted water.
Udvikling af bestande kan indledes med en enkelt hun, eftersom hanner udvikler sig i ubefrugtede æg.
Glass or stainless steel containers with a height of 12 to 20 cm can be used as culture vessels.
Generationer overlapper i vid udstrækning.
The water body should be gently aerated (e.g. 2 bubbles per second) via a pasteur pipette positioned approx.
En hun kan leve mindst 100 dage og kan lægge ca. 100 æg i sin levetid.
2 cm above the sediment surface.
Ved en dyrkningstemperatur på 20 °C opnås dette ved at
To avoid accumulation e.g. of ammonia, the overlying water should be exchanged using a flow-through system, or, at least once a week, manually.
overføre hunner til en ren dyrkningsbeholder og tilsætte tilstrækkeligt foder
The oligochaetes can be held at room temperature with a photo period of 16 hours light (intensity 100 – 1000 lx) and 8 hours dark.
tillade 2-3 dages æglægning og fjerne hunnerne
In the semi-static culture (water renewal once per week), the worms are fed with TetraMin twice a week (e.g. 0,6 - 0,8 mg per cm2 of sediment surface), which can be applied as a suspension of 50 mg TetraMin per ml de-ionized water.
udtage voksne hunner til testen mellem den 28. og den 35. dag efter start og anbringe dem i rene dyrkningsbeholdere.
Lumbriculus variegatus can be removed from the cultures e.g. by transferring substrate with a fine mesh net, or organisms using a fire polished wide mouth (approximately 5 mm diameter) glass pipette, to a separate beaker.
Voksne hunner kan let skelnes fra hanner og andre udviklingstrin, fordi de er større, har en oppustet form og et brunt dorsalt skjold (hanner er tyndere og flade), umodne dyr er hvide til cremefarvede.
If substrate is co-transferred to this beaker, the beaker containing worms and substrate is left overnight under flow-through conditions, which will remove the substrate from the beaker, while the worms remain at the bottom of the vessel.
Midernes udvikling følger omtrent det mønster, der er beskrevet nedenfor, ved 20 °C (figur):
They can then be introduced to newly prepared culture tanks, or processed further for the test as outlined in (3) and (4), or in the following.
Æg 5 dage, larve 2 dage, protonymfe 5 dage, deutonymfe 7 dage, hunnens præ-æglægningsperiode 2 dage.
An issue to be regarded critically when using L. variegatus in sediment tests is its reproduction mode (architomy or morphallaxis, e.g. (6)).
Herefter er miderne voksne.
This asexual reproduction results in two fragments, which do not feed for a certain period until the head or tail part is regenerated (e.g., (7), (8)).
Figur
This means that in L. variegatus exposure via ingestion of contaminated sediment does not take place continuously.
De voksne testdyr fjernes fra den synkroniserede kultur og tilsættes til testbeholderne mellem den 28. og den 35. dag, efter at hunnerne i forældregenerationerne har påbegyndt æglægningen (dvs. 7-14 dage efter at de er blevet voksne).
Therefore, a synchronisation should be performed to minimise uncontrolled reproduction and regeneration, and subsequent high variation in test results.
Det sikrer, at testdyrene allerede har passeret præ-æglægningsperioden og har parret sig med hanner, der også er til stede i dyrkningsbeholderen.
Such variation can occur, when some individuals, which have fragmented and therefore do not feed for a certain time period, are less exposed to the test chemical than other individuals, which do not fragment during the test (9), (10), (11).
Observationer i laboratoriekulturer tyder på, at hunner parrer sig umiddelbart eller kort efter at være blevet voksne, hvis der er hanner til stede (Ruf, Vaninnen, pers. obs.).
10 to 14 days before the start of exposure, the worms should be artificially fragmented (synchronisation).
Perioden på syv dage er valgt for at lette integrationen i laboratorierutinen og mindske variabiliteten i midernes udvikling.
Large (adult) worms, which preferably do not show signs of recent morphallaxis should be selected for synchronisation.
Æglægningen indledes med mindst det antal hunner, der vil være nødvendig til testen (hvis der f.eks. er brug for 400 hunner i testen, lades mindst 400 hunner lægge ægt i 2-3 dage).
These worms can be placed onto a glass slide in a drop of culture water, and dissected in the median body region with a scalpel.
Mindst 1200 æg bør være udgangspunktet for den synkroniserede bestand (kønsforhold ca. 0,5, dødelighed ca. 0,2).
Care should be taken that the posterior ends are of similar size.
For at forhindre kannibalisme er det hensigtsmæssigt ikke at holde mere end 20-30 æglæggende hunner i en beholder.
The posterior ends should then be left to regenerate new heads in a culture vessel containing the same substrate as used in the culture and reconstituted water until the start of exposure.
Ekstraktionsmetoder
Regeneration of new heads is indicated when the synchronised worms are burrowing in the substrate (presence of regenerated heads may be confirmed by inspecting a representative subsample under a binocular microscope).
For mikroartropoder er varmeekstraktion en egnet metode til at udtage dyr fra jorden/substratet (se figur nedenfor).
The test organisms are thereafter expected to be in a similar physiological state.
Metoden er baseret på organismernes aktivitet, hvilket betyder, at kun mobile dyr vil kunne registreres.
This means, that when reproduction by morphallaxis occurs in synchronised worms during the test, virtually all animals are expected to be equally exposed to the spiked sediment.
Princippet i varmeekstraktion er at gøre betingelserne for organismerne gradvist værre i prøven, således at de vil forlade substratet og falde ned i en fikseringsvæske (f.eks. ethanol).
Feeding of the synchronised worms should be done once as soon as the worms are starting to burrow in the substrate, or 7 d after dissection.
Afgørende punkter er ekstraktionens varighed og gradienten for gode til moderate til dårlige betingelser for organismerne.
The feeding regimen should be comparable to the regular cultures, but it may be advisable to feed the synchronised worms with the same food source as is to be used in the test.
Varigheden af ekstraktionen med henblik på økotoksikologiske test skal være så kort som muligt, fordi en eventuel populationstilvækst i ekstraktionsperioden ville give falske resultater.
The worms should be held at test temperature, at 20 ± 2 °C.
På den anden side skal temperatur- og fugtforholdene i prøven altid ligge inden for et interval, der tillader miderne at bevæge sig.
After regenerating, intact complete worms, which are actively swimming or crawling upon a gentle mechanical stimulus, should be used for the test.
Opvarmningen af en jordprøve fører til udtørring af substratet.
Injuries or autotomy in the worms should be prevented, e.g. by using pipettes with fire polished edges, or stainless steel dental picks for handling these worms.
Hvis udtørringen foregår for hurtigt, vil nogle af miderne måske også tørre ud, inden de kan slippe ud.
Sources of starter cultures for Lumbriculus variegatus (addresses in the U.S. adopted from (4))
Tag det hætteglas, der blev brugt til testen.
U.S.A.
Fjern låget, og dæk åbningen med et stykke net eller stof.
Department of Fisheries and Wildlife
Stoffet bør have en maskestørrelse på 1,0-1,5 mm.
No. 13 Natural Resources Building
Fastgør stoffet med en elastik.
Summary of the ring test results
Vend forsigtigt hætteglasset på hovedet, og anbring det i ekstraktionsapparatet.
“Sediment Toxicity Test with Lumbriculus variegatus”
Stoffet forhindrer substrat i at løbe ned i fikseringsvæsken, men tillader mider at forlade prøven.
Results of individual ring test runs:
Påbegynd opvarmningen, når alle hætteglas er anbragt.
Mean worm numbers in the controls and solvent controls at the end of the test;
Afslut ekstraktionen efter 48 timer.
SD = standard deviation;
Fjern hætteglassene med fikseringsvæske, og tæl miderne ved hjælp af et dissektionsmikroskop.
CV = coefficient of variation
Den valgte metodes ekstraktionseffektivitet skal være påvist mindst en eller to gange om året ved hjælp af beholdere, som indeholder et kendt antal unger og voksne mider i ubehandlet testsubstrat.
mean worm number in the controls
Effektiviteten skal gennemsnitligt være ≥ 90 % for alle udviklingstrin samlet set.
mean worm number in the solvent controls
Tullgren-ekstraktionsanordning
interlaboratory mean
Wine mesh barrier
Mean total dry weights of worms per replicate in the controls and solvent controls at the end of the test;
Klargøring af testhætteglasset efter afslutning af testen og inden ekstraktion
CV = coeff. of variation
Identifikation af Hypoaspis (Geolaelaps) aculeifer
total dry weight of worms per replicate (controls)
Underklasse/orden/underorden:
total dry weight of worms per replicate (solvent controls)
Familie:
Toxicity of PCP:
Slægt/underslægt/art:
Summary of endpoints in the ring test;
Forfatter og dato:
interlaboratory means for EC50, NOEC and LOEC;
Deterministiske egenskaber:
minimum detectable difference from the control values during hypothesis testing;
Tectum med afrundet tandkant;
used as a measure of statistical power.
hypostomale riller med mere end seks dentikler;
biological parameter
bageste dorsale setae på Z4 ikke særlig lange;
Inter- laboratory mean (mg/kg)
børsteformede dorsale setae;
Inter- laboratory factor
genitalt skjold normalt, ikke særlig forstørret og når ikke det anale skjold;
geometr. mean (mg/kg)
bageste halvdel af dorsalt skjold uden uparrede setae;
total number of worms
ben II og IV med tykke makrosetae;
total dry weight of worms
dorsal seta Z5 ca. to gange længere end J5;
mortality/survival
fast led på chelicera med 12-14 tænder og bevægeligt led med to tænder;
reproduction (increase of number of worms per replicate)
idiosoma 520-685 μm lang.
growth (biomass increase per replicate)
Hypoaspis miles bruges også til biologisk kontrol og kan forveksles med H. aculeifer.
C.36 PREDATORY MITE (HYPOASPIS (GEOLAELAPS) ACULEIFER) REPRODUCTION TEST IN SOIL
Den største forskel er:
This test method is equivalent to OECD test guideline (TG) 226 (2008).
H. miles tilhører underslægten Cosmolaelaps og har knivlignende dorsale setae, mens H. aculeifer tilhører underslægten Geolaelaps og har børsteformede dorsale setae.
This test method is designed to be used for assessing the effects of chemicals in soil on the reproductive output of the soil mite species Hypoaspis (Geolaelaps) aculeifer Canestrini (Acari: Laelapidae), hence allowing for the estimation of the inhibition of the specific population growth rate (1,2).
Grundlæggende oplysninger om biologien for Hypoaspis (Geolaelaps) aculeifer
Reproductive output here means the number of juveniles at the end of the testing period.
Hypoaspis aculeifer tilhører familien Lealapidae, ordenen Acari (mider), klassen Arachnida, rækken Arthropoda.
H. aculeifer represents an additional trophic level to the species for which test methods are already available.
De lever i alle typer jord og æder andre mider, nematoder, enchytraeider og collemboler (26).
A reproduction test without discrimination and quantification of the different stages of the reproductive cycle is considered adequate for the purpose of this test method.
Ved fødemangel begynder de at æde hinanden (27).
For chemicalsubstances with another exposure scenario than via the soil other approaches might be appropriate (3).
Rovmider består af idiosoma og gnathosoma.
Hypoaspis (Geolaelaps) aculeifer is considered to be a relevant representative of soil fauna and predatory mites in particular.
For så vidt angår idiosoma, kan der ikke klart skelnes mellem prosoma (forkrop) og opisthosoma (abdomen).
It is worldwide distributed (5) and can easily be collected and reared in the laboratory.
Gnathosoma (hovedskjoldet) omfatter instrumenterne til indtagelse af føde, f.eks. palper og chelicera.
A summary on the biology of H. aculeifer is provided in Appendix 7.
Chelicererne er tredelte og forsynet med tænder af forskellig størrelse.
Background information on the ecology of the mite species and the use in ecotoxicological testing is available (4), (5), (6), (7), (8), (9), (10), (11), (12).
Ud over indtagelse af føde bruger hannerne primært deres chelicerer til at overføre spermatoforerne til hunnerne.
Adult females are exposed to a range of concentrations of the test chemical mixed into the soil.
Et dorsalt figur 1skjold dækker idiosoma næsten helt.
The test is started with 10 adult females per replicate vessel.
Kønsorganerne udgør en stor del af idiosoma hos hunner, og de er særlig fremtrædende kort før æglægning.
Males are not introduced in the test, because experience has shown that females mate immediately or shortly after hatching from the deutonymph stage, if males are present.
Der er to ventrale skjold, det sternale skjold og det genitale skjold.
In addition, inclusion of males would prolong the test in a way that the demanding discrimination of age stages would become necessary.
Alle ben er forsynet med børster og torne.
Thus, mating itself is not part of the test.
Børsterne bruges til forankring, når dyret bevæger sig i eller på jorden.
The females are introduced into the test 28-35 days after the start of the egg laying period in the synchronisation (see Appendix 4), as the females can then be considered as already mated and having passed the pre-oviposition stage.
Det første benpar bruges primært som antenner.
At 20 °C the test ends at day 14 after introducing the females (day 0), which allows the first control offspring to reach the deutonymph stage (see Appendix 4).
Det andet benpar bruger miden ikke kun til at bevæge sig med, men også til at fastholde bytte.
For the main measured variable, the number of juveniles per test vessels and additionally the number of surviving females are determined.
Tornene på det fjerde benpar kan benyttes til beskyttelse og som »motor« (28).
The reproductive output of the mites exposed to the test chemical is compared to that of the controls in order to determine the ECx (e.g. EC10, EC50) or the no observed effect concentration (NOEC) (see Appendix 1 for definitions), depending on the experimental design (see paragraph 29).
Hanner er 0,55-0,65 mm lange og vejer 10-15 μg.
An overview of the test schedule is given in Appendix 8.
Hunner er 0,8-0,9 mm lange og vejer 50-60 μg (8)(28) (figur 1).
The water solubility, the log Kow, the soil water partition coefficient and the vapour pressure of the test chemical should preferably be known.
Figur 1
Additional information on the fate of the test chemical in soil, such as the rates of biotic and abiotic degradation, is desirable.
Hun, han, protonymfe og larve af H. aculeifer.
The following criteria should be satisfied in the untreated controls for a test result to be considered valid:
Ved 23 °C bliver miderne kønsmodne efter henholdsvis 16 dage (hunner) og 18 dage (hanner) (6).
Mean adult female mortality should not exceed 20 % at the end of the test;
Hunnerne overfører sperma med solenostom, hvorfra de overføres til ovariet.
The mean number of juveniles per replicate (with 10 adult females introduced) should be at least 50 at the end of the test;
I ovariet modnes og opbevares sperma.
The coefficient of variation calculated for the number of juvenile mites per replicate should not be higher than 30 % at the end of the definitive test.
Befrugtning finder først sted efter modning af sperma i ovariet.
The ECx and/or NOEC of a reference chemical must be determined to provide assurance that the laboratory test conditions are adequate and to verify that the response of the test organisms did not change over time.
De befrugtede eller ubefrugtede æg lægges af hunnerne i klumper eller enkeltvis, fortrinsvis i sprækker eller huller.
Dimethoate (CAS 60-51-5) is a suitable reference chemical that has shown to affect population size (4).
Parrede hunner kan bære afkom af begge køn, hvorimod der fra æg lagt af uparrede hunner kun klækkes afkom af hankøn.
Boric acid (CAS 10043-35-3) may be used as an alternative reference chemical.
Under udviklingen til voksen gennemgås fire udviklingsstadier (æg — larve, larve — protonymfe, protonymfe — deutonymfe, deutonymfe — voksen).
Less experience has been gained with this chemical.
Ægget er mælkehvidt, glasklart, elliptisk og ca. 0,37 mm langt med en fast skal.
Two design options are possible:
Ifølge (8) er larverne 0,42-0,45 mm lange.
The reference chemical can be tested in parallel to the determination of the toxicity of each test chemical at one concentration, which has to be demonstrated beforehand in a dose response study to result in an effect of > 50 % reduction of offspring.
De har kun tre benpar.
In this case, the number of replicates should be the same as that in the controls (see paragraph 29).
I hovedområdet er der udviklet palper og chelicerer.
Alternatively, the reference chemical is tested 1 - 2 times a year in a dose-response test.
Chelicererne, som har nogle få små dentikler, bruges til klækning fra ægget.
Depending on the design chosen, the number of concentrations and replicates and the spacing factor differ (see paragraph 29), but a response of 10 - 90 % effect should be achieved (spacing factor of 1,8).
Efter den første afstødning af larvehud, 1-2 dage efter klækningen, udvikles protonymferne.
The EC50 for dimethoate based on the number of juveniles should fall in the range between 3,0 and 7,0 mg a.s./kg soil (dw).
De er også hvide, 0,45-0,62 mm lange (8) og har fire benpar.
Based on the results obtained with boric acid so far, the EC50 based on the number of juveniles should fall in the range between 100 and 500 mg/kg dw soil.
Tænderne er fuldt udviklet på chelicererne.
Test vessels and equipment
Chelicererne kan også bruges til at rive større partikler ud af klumper af føde (28).
Screw lids are preferred and in that case, the vessels could be aerated twice a week.
Efter endnu en afstødning af hud udvikles deutonymferne.
Alternatively, covers that permit direct gaseous exchange between the substrate and the atmosphere (e.g. gauze) can be used.
De er 0,60-0,80 mm (8) lange og gule til lysebrune.
Since moisture content must be kept high enough during the test, it is essential to control the weight of each experimental vessel during the test and replenish water if necessary.
Fra dette stadium kan der skelnes mellem hunner og hanner.
This may be especially important if no screw lids are available.
Efter en periode med yderligere afstødning af hud, hvor dyrene er inaktive, og det brune skjold udvikler sig (efter ca. 14 dage), er miderne voksne (28)(29)(30).
If a non-transparent test vessel is used, the cover should be made of material that allows for access to light (e.g. by means of a perforated transparent cover) whilst preventing the mites from escaping.
Deres levetid er 48-100 dage ved 25 °C (27).
The size and type of the test vessel depends on the extraction method (see Appendix 5 for details).
Tillæg 8
If heat extraction is applied directly to the test vessel, then a bottom mesh of appropriate mesh size could be added (sealed until extraction), and soil depth should be sufficient to allow for a temperature and moisture gradient.
Sammenfatning og tidsplan for de vigtigste trin i hypoaspistesten
Standard laboratory equipment is required, specifically the following:
Tid (dage)
preferably glass vessels with screw lids;
testens start = dag 0
brushes for transferring mites
Aktivitet/opgave
pH-meter and luxmeter;
til – 28
temperature-controlled incubator or small room;
Overfør hunner fra stamkultur til rene beholdere for at påbegynde synkroniseringen
equipment for extraction (see Appendix 5) (13)
2 dage senere:
overhead light panel with light control
fjernelse af hunner
collection jars for extracted mites.
to eller tre gange om ugen:
For this test, an artificial soil is used.
Giv tilstrækkeligt foder
The artificial soil consists of the following components (all values based on dry mass):
Dag – 5 (+/– 2)
5 % sphagnum peat, air-dried and finely ground (a particle size of 2 ± 1 mm is acceptable);
Klargør syntetisk jord
approximately 74 % air-dried industrial sand (depending on the amount of CaCO3 needed), predominantly fine sand with more than 50 % of the particles between 50 and 200 microns.
Dag – 4 (+/– 2)
The exact amount of sand depends on the amount of CaCO3 (see below), together they should add up to 75 %.
Bestem den syntetiske jords WHC
< 1,0 % calcium carbonate (CaCO3, pulverised, analytical grade) to obtain a pH of 6,0 ± 0,5;
Tør natten over
the amount of calcium carbonate to be added may depend principally on the quality/nature of the peat (see Note 1).
Næste dag:
The amount of CaCO3 required will depend on the components of the soil substrate and should be determined by measuring the pH of soil sub-samples immediately before the test (14).
Vej prøver, og beregn WHC
The peat content of the artificial soil deviates from other test methods on soil organisms, where in most cases 10 % peat is used (e.g. (15)).
Dag – 4 (+/– 2)
However, according to EPPO (16) a typical agricultural soil has not more than 5 % organic matter, and the reduction in peat content thus reflects the decreased possibilities of a natural soil for sorption of the test chemical to organic carbon.
Fugt syntetisk jord til en WHC på 20-30 %
If required, e.g. for specific testing purposes, natural soils from unpolluted sites may also serve as test and/or culture substrate.
Start testen:
However, if natural soil is used, it should be characterised at least by origin (collection site), pH, texture (particle size distribution) and organic matter content.
Tilsæt testkemikalie til den syntetiske jord
If available, the type and name of the soil according to soil classification should be included, and the soil should be free from any contamination.
Tilsæt ti hunner til hver replikat
In case the test chemical is a metal or organo-metal, the cation exchange capacity (CEC) of the natural soil should also be determined.
Vej hver replikat
Special attention should be paid to meet the validity criteria as background information on natural soils typically is rare.
Opstil abiotiske kontroller for fugtindhold og pH, to replikater for hver behandling
For the determination of pH a mixture of soil and 1 M potassium chloride (KCl) or 0,01 M calcium chloride (CaCl2) solution in a 1:5 ratio is used (see (14) and Appendix 3).