natgillin/translations-raw huggingface dataset:
Rapportering af testresultater
Females are 0,8 - 0,9 mm long and are weighing 50 - 60 μg (8) (28) (Fig 1).
Undersøgelsesdataene skal omfatte:
Figure 1
Testlaboratorium:
Female, male, protonymph and larvae of H. aculeifer.
Ansvarligt personale og deres undersøgelsesansvar
At 23 °C, the mites become sexually mature after 16 days (females) and 18 days (males), respectively (6).
Hvert laboratorium skal have dokumenteret kendskab til en række repræsentative kemikalier.
The females carry over the sperms by the solenostom where they will be then transferred to the ovar.
Karakterisering af testkemikalie
In the ovar the sperms mature and will be stored.
Fysisk form og relevante fysisk-kemiske egenskaber
Fertilisation takes place only after maturation of the sperms in the ovar.
Klargøring af testkoncentrationer: metode og frekvens
The fertilised or unfertilised eggs will be deposited by the females in clumps or separately, preferably in crevices or holes.
Information om stabilitet og bionedbrydelighed.
Copulated females can bear juveniles of both sexes whereas from eggs of uncopulated females only male juveniles are hatching.
Opløsningsmiddel:
During development to the adult four phases of development (egg — larvae, larvae — protonymph, protonymph — deutonymph, deutonymph — adult) are passed through.
Karakterisering af opløsningsmiddel (form, anvendt koncentration)
The egg is milky white, hyaline, elliptical and approximately 0,37 mm long with a solid mantle.
Begrundelse for valg af opløsningsmiddel (hvis andet end vand).
According to (8), the larvae are between 0,42 - 0,45 mm in size.
Testdyr:
They have only three pairs of legs.
Art og stamme.
In the head region palps and chelicers are developed.
Leverandør og særlig leverandør
The chelicers, having some few small denticles, are used to hatch from the egg.
Fiskens alder ved begyndelsen af testen og reproduktiv tilstand/gydningstilstand
After the first moult, 1 - 2 days after hatching, the protonymphs are developed.
Oplysninger om procedure for akklimatisering af dyr
They are also white, the size is 0,45 - 0,62 mm (8) and they have four pairs of legs.
Fiskenes kropsvægt ved eksponeringsperiodens start (fra en delprøve af fiskebestanden).
On the chelicers the teeth are completely present.
Testbetingelser:
Beginning with that stadium the mites start to forage.
Anvendt testprocedure (testtype, belastningsgrad, bestandtæthed osv.)
For that reason the cuticula of the prey is pierced with the chelicers and a secretion for the extra intestinal digestion is emitted into the prey.
Metode til fremstilling af stamopløsning og gennemstrømningshastighed
The food mash can then be sucked by the mite.
De nominelle testkoncentrationer, ugentligt målte koncentrationer af testopløsningerne og anvendte analysemetoder, de målte værdiers gennemsnit og standardafvigelser i testbeholderne samt dokumentation for, at målingerne refererer til testkemikaliets faktiske koncentration i opløsning
The chelicers can also be used to rip bigger particles out of food nuggets (28).
Fortyndingsvandets karakteristika (herunder pH, hårdhedsgrad, alkalinitet, temperatur, koncentration af opløst ilt, restindhold af chlor, samlet mængde organisk kulstof, opslæmmet tørstof og samtlige andre udførte målinger)
After one further moult the deutonymphs are developed.
Vandkvalitet i testbeholderne:
They are 0,60 - 0,80 mm (8) in size and yellow to light brown in colour.
pH, hårdhedsgrad, temperatur og koncentration af opløst ilt
Beginning with that phase they can be separated into females and males.
Detaljerede oplysninger om fodring (f.eks. foderets type og kilde samt fodermængde og fodringshyppighed såvel som analyser for relevante kontaminanter, hvis de foreligger (f.eks. PCB'er, PAH'er og organochlorpesticider).
After further ecdysis, during which time the animals are inactive and the brown shield is developing (approx. after 14 days) the mites are adult (28) (29) (30).Their life span is between 48 and 100 days at 25 °C (27).
Bevis for, at kontrollerne opfylder testens acceptkriterier
Appendix 8
Data om eventuel dødelighed ved enhver testkoncentration og kontrol
Summary and time schedule of the main actions to be taken in order to perform the Hypoaspis test
Anvendte statistiske analyseteknikker, behandling af data og begrundelse for de anvendte teknikker
Time (days)
Data om biologiske observationer af makromorfologi, herunder sekundære kønskarakteristika og vitellogenin
test start = day 0
Resultater af dataanalyserne, fortrinsvis i tabelform og grafisk form
Activity / task
Forekomst af eventuelle usædvanlige reaktioner hos fiskene samt eventuelle synlige virkninger frembragt af testkemikaliet.
Day – 35
VEJLEDNING I FORTOLKNING OG GODKENDELSE AF TESTRESULTATER
to – 28
Denne del beskriver overvejelser, der bør tages i betragtning i forbindelse med fortolkningen af testresultaterne for de forskellige målte endepunkter.
Transfer females from stock culture to clean vessels to start synchronisation
Resultaterne bør fortolkes med forsigtighed, når testkemikaliet tilsyneladende forårsager åbenlys toksicitet eller har indvirkning på testdyrets generelle tilstand.
2 days later:
Ved fastsættelse af testkoncentrationsintervallet skal det sikres, at den maksimalt tolerede koncentration ikke overstiges, for at give mulighed for en relevant fortolkning af dataene.
removal of females
Det er vigtigt at have mindst en behandling, hvor der ikke er nogen tegn på toksiske virkninger.
Twice or three times a week:
Tegn på sygdom og tegn på toksiske virkninger bør vurderes nøje og anføres i rapporten.
supply with sufficient food
Det er f.eks. muligt, at produktion af VTG hos hunner også kan påvirkes af generel toksicitet og ikke-hormonforstyrrende toksiske virkemåder, f.eks. levertoksicitet.
Day – 5 (+/- 2)
Fortolkningen af virkningerne kan dog styrkes af andre behandlingsniveauer, som ikke konfunderes af systemisk toksicitet.
Prepare artificial soil
Nogle få aspekter skal overvejes, hvis testresultaterne skal godkendes.
Day – 4 (+/- 2)
Som hovedregel skal VTG-niveauerne måles hos kontrolgrupper af hanner og hunner hver for sig og være adskilt med omkring tre størrelsesordener hos tykhovedet elritse og zebrafisk og omkring en størrelsesorden hos japansk risfisk.
Determine WHC of artificial soil
Eksempler på værdiintervallerne i kontrol- og behandlingsgrupperne findes i valideringsrapporterne (1, 2, 3, 4).
Dry over night
Høje VTG-værdier hos hanner i kontrollerne kan kompromittere assayets respons og evne til at påvise svage østrogenagonister.
Next day:
Lave VTG-værdier hos hunner i kontrollerne kan kompromittere assayets respons og evne til at påvise aromataseinhibitorer og østrogenagonister.
weigh samples and calculate WHC
Valideringsundersøgelserne dannede udgangspunkt for vejledningen.
Day – 4 (+/– 2)
Hvis et laboratorium ikke har udført assayet før, eller der er foretaget væsentlige ændringer (f.eks. ny fiskestamme eller leverandør) anbefales det at undersøge dets tekniske kompetence.
Pre moisture artificial soil to achieve 20 - 30 % of WHC
Det anbefales at bruge kemikalier med en række virkemåder eller indvirkning på en række testendepunkter.
Start test:
I praksis opfordres hvert laboratorium til at opbygge sine egne historiske kontroldata for hanner og hunner og teste et positivt kontrolkemikalie for østrogenaktivitet (f.eks. østradiol-17β ved 100 ng/l eller en kendt svag agonist), som fører til øget VTG hos hanfisk, et positivt kontrolkemikalie for aromataseinhibering (f.eks. fadrozol eller prochloraz ved 300 μg/l), som fører til reduceret VTG hos hunfisk, og et positivt kontrolkemikalie for androgen aktivitet (f.eks. trenbolon-17β ved 5 μg/l), som fører til induktion af sekundære kønskarakteristika hos tykhovedet elritse- og japansk risfisk-hunner.
add test chemical to artificial soil
Alle disse data kan sammenlignes med tilgængelige data fra valideringsundersøgelserne (1, 2, 3), for at sikre at laboratoriet har den fornødne tekniske kompetence.
Introduce 10 females to each replicate
Generelt betragtes VTG-målinger som positive, hvis der er en statistisk signifikant stigning i VTG hos hanner (p < 0,05) eller et statistisk signifikant fald hos hunner (p < 0,05), som minimum ved den højeste testede dosis, sammenlignet med kontrolgruppen, og hvis der ikke er tegn på generel toksicitet.
Weigh each replicate
Et positivt resultat underbygges yderligere af påvisningen af et biologisk sandsynligt forhold mellem dosis- og responskurven.
Set up abiotic controls for moisture content and pH, 2 replicates for each treatment
Som nævnt tidligere har faldet i vitellogenin muligvis ikke udelukkende endokrin oprindelse;
Dry moisture controls over night
et positivt resultat bør dog generelt fortolkes som bevis for hormonforstyrrende virkning in vivo og bør normalt give anledning til yderligere afklaring.
weigh moisture controls
Forkortelser og definitioner
measure pH of dried abiotic controls
Kemikalie
Day 3, 6, 9, 12 (approx.)
et stof eller en blanding.
Supply each replicate with sufficient amount of prey organisms
Variationskoefficient.
Weigh each replicate and eventually add evaporated water
Enzymkoblet immunadsorptionsteknik.
Terminate test, set up extraction with all replicates plus extraction efficiency controls
Belastningsgrad
Dry water content controls over night
Fiskens vådvægt pr. vandvolumen.
weigh water content controls
Bestandtæthed
measure pH of dried controls
Antal fisk pr. vandvolumen.
Day 16
Phospholipid-glycoprotein-precursor for æggeblommeprotein, der normalt forekommer hos seksuelt aktive hunner af alle æglæggende arter.
Terminate extraction
HPG-aksen
Day 16 +
Hypothalamus-hypofyse-gonadeakse.
Record number of adults and juveniles in extracted material
Maksimalt tolereret koncentration, som udgør ca. 10 % af LC50.
Report results on template tables
Testbetingelser for assay til screening af det endokrine system hos fisk
Report testing procedure in test protocol sheets
Anbefalet art
C.37.
Tykhovedet elritse
AY FISH ASSAY:
Zebrafisk
A SHORT-TERM SCREENING FOR OESTROGENIC AND ANDROGENIC ACTIVITY, AND AROMATASE INHIBITION
Testtype
This test method is equivalent to OECD test guideline (TG) 230 (2009).
Gennemstrømning
The need to develop and validate a fish assay capable of detecting certain endocrine active chemicals originates from the concerns that environmental levels of chemicals may cause adverse effects in both humans and wildlife due to the interaction of these chemicals with the endocrine system.
Vandtemperatur
In 1998, the OECD initiated a high-priority activity to revise existing guidelines and to develop new guidelines for the screening and testing of potential endocrine disrupters.
Belysningskvalitet
One element of the activity was to develop a Test Guideline for the screening of chemicals active on the endocrine system of fish species.
Fluorescerende pærer (bredspektret)
The 21-day Fish Endocrine Screening Assay underwent an extensive validation programme consisting of inter-laboratory studies with selected chemicals to demonstrate the relevance and reliability of the assay for the detection of oestrogenic and aromatase inhibiting chemicals (1, 2, 3, 4, 5) in the three fish species investigated (the fathead minnow, the Japanese medaka and the zebrafish);
10-20 μE/m2/s, 540-1000 lux eller 50-100 ft-c (rumniveauer i laboratoriet)
the detection of androgenic activity is possible in the fathead minnow and the medaka, but not in the zebrafish.
Lysperiode (daggry/skumringsovergange er valgfri, men betragtes ikke som nødvendige)
This test method does not allow the detection of anti-androgenic chemicals.
16 t. lys, 8 t. mørke
The validation work has been peer-reviewed by a panel of experts nominated by the National Coordinators of the Test Guideline Programme (6).
12-16 t. lys, 12-8 t. mørke
The assay is not designed to identify specific mechanisms of hormonal disruption because the test animals possess an intact hypothalamic-pituitary-gonadal (HPG) axis, which may respond to chemicals that impact on the HPG axis at different levels.
< 5 g/l
The Fish Short Term Reproduction assay (OECD TG 229) includes fecundity and, as appropriate, gonadal histopathology for the fathead minnow, as well as all endpoints included in this test method.
Testkammerstørrelse
OECD TG 229 provides a screening of chemicals which affect reproduction through various mechanisms including endocrine modalities.
Volumen testopløsning
This should be considered prior to selecting the most appropriate test method.
1,5 l (minimum)
This test method describes an in vivo screening assay where sexually mature male and spawning female fish are held together and exposed to a chemical during a limited part of their life-cycle (21 days).
Volumenudskiftning af testopløsninger
At termination of the 21-day exposure period, depending on the species used, one or two biomarker endpoint(s) are measured in males and females as indicators of oestrogenic, aromatase inhibition or androgenic activity of the test chemical;
Mindst 6 dagligt
these endpoints are vitellogenin and secondary sexual characteristics.
Mindst 5 dagligt
Vitellogenin is measured in fathead minnow, Japanese medaka and zebrafish, whereas secondary sex characteristics are measured in fathead minnow and Japanese medaka only.
Se punkt 20.
This bioassay serves as an in vivo screening assay for certain endocrine modes of action and its application should be seen in the context of the “OECD Conceptual Framework for the Testing and Assessment of Endocrine Disrupting Chemicals” (28).
Hanner:
INITIAL CONSIDERATIONS AND LIMITATIONS
Hunner:
Vitellogenin is normally produced by the liver of female oviparous vertebrates in response to circulating endogenous oestrogen.
Antal fisk pr. testbeholder
It is a precursor of egg yolk proteins and, once produced in the liver, travels in the bloodstream to the ovary, where it is taken up and modified by developing eggs.
6 (2 hanner og 4 hunner)
Vitellogenin is almost undetectable in the plasma of immature female and male fish because they lack sufficient circulating oestrogen;
10 (5 hanner og 5 hunner)
however, the liver is capable of synthesizing and secreting vitellogenin in response to exogenous oestrogen stimulation.
Antal behandlinger
The measurement of vitellogenin serves for the detection of chemicals with various oestrogenic modes of action.
= 3 (plus passende kontroller)
The detection of oestrogenic chemicals is possible via the measurement of vitellogenin induction in male fish, and it has been abundantly documented in the scientific peer-reviewed literature (e.g. (7)).
Antal beholdere pr. behandling
Vitellogenin induction has also been demonstrated following exposure to aromatizable androgens (8, 9).
Minimum 4
A reduction in the circulating level of oestrogen in females, for instance through the inhibition of the aromatase converting the endogenous androgen to the natural oestrogen 17β-estradiol, causes a decrease in the vitellogenin level, which is used to detect chemicals having aromatase inhibiting properties (10, 11).
Minimum 2
The biological relevance of the vitellogenin response following oestrogenic/aromatase inhibition is established and has been broadly documented.
Antal fisk pr. testkoncentration
However, it is possible that production of VTG in females can also be affected by general toxicity and non-endocrine toxic modes of action, e.g. hepatotoxicity.
16 voksne hunner og 8 hanner (4 hunner og 2 hanner i hver replikatbeholder)
Several measurement methods have been successfully developed and standardised for routine use.
10 voksne hunner og 10 hanner (5 hunner og 5 hanner i hver replikatbeholder)
This is the case of species-specific Enzyme-Linked Immunosorbent Assay (ELISA) methods using immunochemistry for the quantification of vitellogenin produced in small blood or liver samples collected from individual fish (12, 13, 14, 15, 16, 17, 18).
Saltsøkrebs, nauplii, to eller tre gange om dagen (ad libitum), kommercielt tilgængeligt foder eller en kombination af ovenstående
Appendix 6 provides the recommended procedures for sample collection for vitellogenin analysis.
Ingen, medmindre DO-koncentration falder under 60 % luftmætning
Kits for the measurement of vitellogenin are widely available;
Rent overflade- eller brøndvand, rekonstitueret vand eller afkloret ledningsvand
such kits should be based on a validated species-specific ELISA method.
Præ-eksponeringsperiode
Secondary sex characteristics in male fish of certain species are externally visible, quantifiable and responsive to circulating levels of endogenous androgens;
7 dage anbefales
this is the case for the fathead minnow and the medaka — but not for zebrafish, which does not possess quantifiable secondary sex characteristics.
Varighed af eksponering for kemikaliet
Females maintain the capacity to develop male secondary sex characteristics, when they are exposed to androgenic chemicals in water.
Biologiske endepunkter
Several studies are available in the scientific literature to document this type of response in fathead minnow (20) and medaka (21).
overlevelse
A decrease in secondary sex characteristics in males should be interpreted with caution because of low statistical power, and should be based on expert judgement and weight of evidence.
adfærd
There are limitations to the use of zebrafish in this assay, due to the absence of quantifiable secondary sex characteristics responsive to androgenic acting chemicals.
sek. kønskarakteristika
In the fathead minnow, the main indicator of exogenous androgenic exposure is the number of nuptial tubercles located on the snout of the female fish.
Testgodkendelse
In the medaka, the number of papillary processes constitutes the main marker of exogenous exposure to androgenic chemicals in female fish.
Opløst ilt > 60 % af mætning;
Appendix 5A and Appendix 5B indicate the recommended procedures to follow for the evaluation of sex characteristics in fathead minnow and in medaka, respectively.
gennemsnitlig temperatur på 25 °C ± 2 °C;
Definitions used in this test method are given in Appendix 1.
90 % overlevelse for fisk i kontrollerne;
In the assay, male and female fish in a reproductive status are exposed together in test vessels.
målte testkoncentrationer inden for 20 % af de gennemsnitlige målte værdier pr. behandlingsniveau.
Their adult and reproductive status enables a clear differentiation of each sex, and thus a sex-related analysis of each endpoint, and ensures their sensitivity towards exogenous chemicals.
gennemsnitlig temperatur på 24 °C ± 2 °C;
At test termination, sex is confirmed by macroscopic examination of the gonads following ventral opening of the abdomen with scissors.
gennemsnitlig temperatur på 26 °C ± 2 °C;
An overview of the relevant bioassay conditions is provided in Appendix 2.
Kemiske egenskaber for acceptabelt fortyndingsvand
The assay is normally initiated with fish sampled from a population that is in spawning condition;
< 20 mg/l
senescent animals should not be used.
< 2 mg/l
Guidance on the age of fish and on the reproductive status is provided in the section on Selection of fish.
Total organiske phosphorpesticider
The assay is conducted using three chemical exposure concentrations as well as a water control, and a solvent control if necessary.
Tillæg 4A
Two vessels or replicates per treatment are used (each vessel containing 5 males and 5 females) in medaka and zebrafish, whereas four vessels or replicates per treatment are used (each vessel containing 2 males and 4 females) in fathead minnow.
Gydningssubstrat til zebrafisk
This is to accommodate the territorial behaviour of male fathead minnow while maintaining sufficient power of the assay.
Gydningsbakke
The exposure is conducted for 21 days and sampling of fish is performed at day 21 of exposure.
instrumentbakke af glas, f.eks. 22 × 15 × 5,5 cm (l × b × d), dækket med et aftageligt rustfrit metalgitter (maskebredde 2 mm).
On sampling at day 21, all animals are killed humanely.
Gitteret bør dække instrumentbakkens åbning på et niveau under kanten.
Secondary sex characteristics are measured in fathead minnow and medaka (see Appendix 5A and Appendix 5B);
Gydningssubstratet fastgøres på gitteret.
blood samples are collected for determination of vitellogenin in zebrafish and fathead minnow, alternatively head/tail can be collected for the determination of vitellogenin in zebrafish (Appendix 6);
Plastmaterialet bør være udvasket i et tilstrækkeligt volumen varmt vand, til at sikre at ingen kemikalier kan afgives til testvandet.
liver is collected for VTG analysis in medaka (Appendix 6).
Ved brug af glasmaterialer sikres det, at fiskene hverken skades eller sidder fast i forbindelse med deres kraftige aktivitet.
TEST ACCEPTANCE CRITERIA
Afstanden mellem bakken og glaspladen bør være mindst 3 cm, for at sikre at gydningen ikke sker uden for bakken.
For the test results to be acceptable the following conditions apply:
De æg, der lægges på bakken, falder gennem gitteret og kan udtages 45-60 min. efter belysningens start.
the mortality in the water (or solvent) controls should not exceed 10 % at the end of the exposure period;
De gennemsigtige æg er ikke-klæbende og kan let tælles med tværgående lys.
the dissolved oxygen concentration should be at least 60 % of the air saturation value (ASV) throughout the exposure period;
Når der bruges fem hunner pr. beholder, kan et antal æg på op til 20 om dagen betragtes som lavt, op til 100 som middel og over 100 som højt.
the water temperature should not differ by more than ± 1,5 °C between test vessels at any one time during the exposure period and be maintained within a range of 2 °C within the temperature ranges specified for the test species (Appendix 2);
Gydningsbakken fjernes, æggene indsamles, og gydningsbakken anbringes i testbeholderen igen, enten så sent som muligt om aftenen eller meget tidligt om morgenen.
evidence should be available to demonstrate that the concentrations of the test chemical in solution have been satisfactorily maintained within ± 20 % of the mean measured values.
Bakken skal anbringes senest efter en time, da gydningssubstratet ellers kan forårsage individuel parring og gydning på et usædvanligt tidspunkt.
Normal laboratory equipment and especially the following:
Hvis situationen kræver, at gydningsbakken anbringes senere, skal det ske mindst ni timer efter belysningens start.
oxygen and pH meters;
På dette sene tidspunkt på dagen induceres gydning ikke længere.
equipment for determination of water hardness and alkalinity;
Tillæg 4B
adequate apparatus for temperature control and preferably continuous monitoring;
Gydningssubstrat til tykhovedet elritse
tanks made of chemically inert material and of a suitable capacity in relation to the recommended loading and stocking density (see Appendix 2);
To eller tre kombinerede gydningsfliser og bakker af plast/keramik/glas eller rustfrit stål anbringes i hvert testkammer (f.eks. 80 mm lang grå halvcirkelformet rende på en 130 mm lang bakke med kanter) (se billede).
spawning substrate for fathead minnow and zebrafish, Appendix 4 gives the necessary details;
Korrekt tørrede PVC-fliser eller keramikfliser har vist sig at være egnede til at modtage et gydningssubstrat (Thorpe et al., 2007).
suitably accurate balance (i.e. accurate to ± 0,5 mg).
Det anbefales, at fliserne slibes for at fremme adhæsion.
Any water in which the test species shows suitable long-term survival and growth may be used as test water.
Bakken bør desuden afskærmes for at hindre fiskenes adgang til de nedfaldne æg, medmindre det har vist sig, at æg klæber til det anvendte gydningssubstrat.
It should be of constant quality during the period of the test.
Bunden er udformet, så den indeholder eventuelle æg, der ikke klæber til flisens overflade og derfor falder ned i bunden af beholderen (eller de æg, der lægges direkte på den flade plastbund).
The pH of the water should be within the range 6,5 to 8,5, but during a given test it should be within a range of ± 0,5 pH units.
Alt gydningssubstrat bør udvaskes i mindst 12 timer i fortyndingsvand inden brug.
In order to ensure that the dilution water will not unduly influence the test result (for example by complexion of test chemical), samples should be taken at intervals for analysis.