natgillin/translations-raw huggingface dataset:
Adequate statistical power to detect a 20 % difference from the control at the 5 % level of significance (α = 0,05) should be demonstrated.
iga 1. ja 2. põlvkonna nõu kohta nende vastsete arv, kellest sääski ei arenenud;
With metric responses (e.g. development rate), the t-test is a suitable statistical method if data meet the requirements of this test (normality, homogeneous variances).
ja 2. põlvkonnas väljaarenemise protsent/osakaal iga paralleelkatse ja kontsentratsiooni kohta (isas- ja emassääsed kokku);
An unequal-variance t-test or a non-parametric test, such as the Wilcoxon-Mann-Whitney test may be used, if these requirements are not fulfilled.
ja 2. põlvkonnas elusate ja väljaarenenud sääskede keskmine arengukiirus igas paralleelkatses ja igal kontsentratsioonil (isas- ja emassääsed eraldi ja ka koos);
With the emergence ratio, Fisher's exact test is appropriate.
kristallisaatorisse munetud munanööride arv paljunduspuuri ja iga päeva kohta;
Preparation of the water-sediment system (water spiking)
iga munanööri kirjeldus (suurus, kuju ja viljakus);
a.
sigivus – munanööride üldarv paljunduspuuridesse pandud emassääskede üldarvu kohta;
Formulated sediment (see paragraphs 13-14 and Appendix 3) is added to each test vessel and crystallising dish to form a layer of at least 1,5 cm (for the crystallising dish it may be somewhat lower) but maximally 3 cm.
viljakus – viljakate munanööride üldarv paljunduspuuridesse pandud emassääskede üldarvu kohta;
Water (see paragraph 15) is added so that the ratio of the depth of the sediment layer and the depth of the water does not exceed 1:4.
mürgisuse näitajate hinnangud, näiteks ECx (ja selle usaldusvahemik), täheldatavat toimet mitteavaldav kontsentratsioon (NOEC) ja selle määramiseks kasutatud statistilised meetodid;
After preparation of the test vessels the sediment-water system should be left under gentle aeration for approximately seven days prior to addition of the first instar larvae of the 1st or 2nd generation (see paragraph 14 and Appendix 3).
Käesoleva lisa peatükk C.28 „Sette-vee mürgisuskatse surusääsklastel rikastatud vee kasutamisega”.
The sediment-water system of the crystallising dishes is not aerated during the test, since they do not need to support larval survival (before hatching the egg ropes are already collected).
Käesoleva lisa peatükk C.27 „Sette-vee mürgisuskatse surusääsklastel rikastatud sette kasutamisega”.
To avoid separation of sediment ingredients and re-suspension of fine material during addition of test water in the water column, the sediment can be covered with a plastic disc while water is poured onto it.
J. Amer.
The disc is removed immediately afterwards.
OECD (2006), Current Approaches in the Statistical Analysis of Ecotoxicity Data: a Guidance to Application, OECD Series on Testing and Assessment No.
Other devices may also be appropriate.
54, 146 pp., ENV/JM/MONO(2006)18, OECD, Paris.
Preparation of the water-sediment system (spiked sediment)
Spetsiaalselt koostatud sete ehk taastatud, tehislik või sünteetiline sete selline materjalide segu, mida kasutatakse loodusliku sette füüsiliste koostisosade imiteerimiseks.
b.
Kattev veekiht vesi, mis on pandud katsenõus sette peale.
The spiked sediments prepared according to paragraph 16b are placed in the vessels and crystallising dish and overlying water is added to produce a sediment-water volume ratio of 1:4.
Poorivesi vesi, mis asub sette- ja pinnaseosakeste vahelises ruumis.
The depth of the sediment layer should be in the range of 1,5 to 3 cm (it may be somewhat lower for the crystallising dish).
Rikastatud vesi katses kasutatav vesi, millele on lisatud uuritav kemikaal.
To avoid separation of sediment ingredients and re-suspension of fine material during addition of test water in the water column, the sediment can be covered with a plastic disc while water is poured onto it, and the disc removed immediately afterwards.
Chironomus riparius'e kasvatamist käsitlevad soovitused
Appropriate equilibration time is sediment and chemical specific, and can be in the order of hours to days and in rare cases up to five weeks.
Mahuti põhi kaetakse ligikaudu 5–10 mm paksuse peene kvartsliiva kihiga.
As this would leave time for degradation of many chemicals, equilibrium is not awaited but an equilibration period of 48 hours is recommended.
Diatomiit (näiteks Merck, Art 8117) on samuti tõendatult sobiv substraat (piisab õhemast kihist, isegi mõnest millimeetrist).
However, when the degradation half-life of the chemical in sediment is known to be long (see paragraph 8), the equilibration time may be extended.
Seejärel lisatakse sobivat vett mitme cm paksuse kihina.
At the end of this further equilibration period, the concentration of the test chemical should be measured in the overlying water, the pore water and the sediment, at least at the highest concentration and a lower one (see paragraph 38).
Vett tuleks vajaduse korral lisada aurumiskadude asendamiseks ja kuivamise vältimiseks.
These analytical determinations of the test chemical allow for calculation of a mass balance and expression of results based on measured concentrations.
Vee võib vajaduse korral välja vahetada.
Test vessels should be covered (e.g. by glass plates).
Tuleks tagada kerge aereerimine.
If necessary, during the study the water levels may be topped up to the original volume in order to compensate for evaporation.
Vastsete kasvatamise nõusid tuleks hoida sobivas puuris, mis takistab väljaarenenud täiskasvanud isendite väljapääsu.
This should be performed using distilled or deionised water to prevent any build-up of salts.
Puur peaks olema piisavalt suur, et võimaldada väljaarenenud valmikute parvedesse kogunemist, vastasel juhul ei pruugi toimuda paljunemist (miinimumsuurus on ligikaudu 30 × 30 × 30 cm).
Crystallising dishes in the breeding cages are not covered and may, but do not need to be adjusted to compensate for water loss during the test period, since the egg ropes are only in contact with the water for about one day and the dishes are only used during a short phase of the test.
Puure tuleks hoida toatemperatuuril või püsiva keskkonnaga ruumis temperatuuril 20 ± 2 °C 16-tunnise valgustusperioodiga (intensiivsus ligikaudu 1000 luksi) ja 8 tunni pimedusega.
Addition of test organisms
On teada, et vähem kui 60 %-line suhteline õhuniiskus võib paljunemist takistada.
Four to five days before adding the first instar larvae for the 1st generation, egg masses should be taken from the culture and placed in small vessels in culture medium.
Võib kasutada mis tahes looduslikku või sünteetilist vett.
Aged medium from the stock culture or freshly prepared medium may be used.
Enamasti kasutatakse kaevuvett, deklooritud kraanivett ja tehislikke kasvukeskkondi (näiteks Elendti kasvukeskkond M4 või M7, vt allpool).
In any case, a small amount of food, e.g. a few droplets of filtrate from a finely ground suspension of flaked fish food, should be added to the culture medium (see Appendix 2).
Vett tuleks enne kasutamist aereerida.
Only freshly laid egg masses should be used.
Vajaduse korral võib kultuuri kasvuvett uuendada, eemaldades kasutatud vee kultuuri kasvunõudest ettevaatlikult valamise teel või sifooniga, kahjustamata vastsete kestasid.
Normally, the larvae begin to hatch a couple of days after the eggs are laid (2 to 3 days for C. riparius at 20 °C and 1 to 4 days for C. dilutus at 23 °C and C. yoshimatsui at 25 °C) and larval growth occurs in four instars, each of 4-8 days duration.
Vastsete söötmine
First instar larvae (maximum 48 h post hatching) should be used in the test.
Chironomus'e vastseid tuleks sööta helbelise kalasöödaga (Tetra Min®, Tetra Phyll® või muu samalaadne kaubanduslik kalasööda tootemark) koguses ligikaudu 250 mg nõu kohta päevas.
The instar stage of larvae can potentially be checked using head capsule width (7).
Sööta võib anda kuiva jahvatatud pulbrina või suspensioonina vees:
Twenty first instar larvae for the 1st generation are allocated randomly to each test vessel containing the sediment-water system, using a blunt pipette.
1,0 g helbelist kalasööta lisatakse 20 ml lahjendusveele ja see segatakse homogeense segu saamiseni.
Aeration of the water is stopped whilst adding larvae to test vessels and should remain so for 24 hours following addition of larvae (see paragraph 32).
Seda preparaati võib sööta koguses 5 ml nõu kohta päevas (loksutage enne kasutamist).
According to the test design used (see paragraphs 19 and 20), the number of larvae used per concentration is at least 120 (6 replicates per concentration) for the ECX approach and 160 for the NOEC approach (8 replicates per concentration).
Vanemad vastsed võivad vajada rohkem sööta.
In the spiked sediment design, exposure starts with the addition of the larvae.
Söötmist kohandatakse vee kvaliteediga.
Spiking the overlying water
Kui kultuuri kasvukeskkond muutub häguseks, tuleks sööda hulka vähendada.
Twenty-four hours after adding the first instar larvae for the 1st generation, the test chemical is spiked into the overlying water column, and slight aeration is again supplied (for possible amendments of the test design, see paragraph 7).
Sööda lisamist tuleks hoolikalt jälgida.
Small volumes of the test chemical stock solutions are applied below the surface of the water using a pipette.
Liiga vähe sööta põhjustab vastsete rände veesamba suunas ning liiga palju sööta hoogustab mikroobide tegevust ja vähendab vee hapnikusisaldust.
The overlying water should then be mixed with care not to disturb the sediment.
Mõlemad tingimused võivad vähendada kasvu kiirust.
In the spiked water design, exposure starts with the spiking of the water (i.e. one day after addition of the larvae).
Uute kultuuri kasvunõude valmisseadmisel võib lisada ka mõne rohevetika (näiteks Scenedesmus subspicatus, Chlorella vulgaris) rakke.
Collecting emerged adults
Väljaarenenud valmikute söötmine
Emerged midges of the 1st generation are collected at least once, but preferably twice a day (see point 36) from the test vessels using an aspirator, exhauster or similar device (see Appendix 5).
Mõned teadlased on soovitanud väljaarenenud valmikute söötmiseks kasutada küllastunud sahharoosilahuses niisutatud puuvillast lappi.
Special care should be taken not to damage the adults.
Temperatuuril 20 ± 2 °C hakkavad ligikaudu 13–15 päeva möödumisel vastsete kasvatamise nõudest väljuma valmikud.
The collected midges from four test vessels within one treatment are released into a breeding cage to which they had been previously assigned.
Isased on tänu sulgjatele tundlatele ja kõhnemale kehale kergesti äratuntavad.
At the day of first (male) emergence, crystallising dishes are spiked by pipetting a small volume of the test chemical stock solution below the water surface (spiked water design).
Munamassid
The concentration of test chemical in the crystallising dish is nominally the same as in the treatment vessels which are assigned to that specific breeding cage.
Kui valmikud on paljunduspuuris, tuleks kõiki vastsete kasvatamise nõusid kontrollida kolm korda nädalas sinna želatiinjate munamasside lisandumise suhtes.
For the spiked sediment design, the crystallising dishes are prepared at around day 11 after the start of the exposure (i.e. addition of the 1st generation larvae) so that they can equilibrate for about 48 hours before the first egg ropes are produced.
Kui leitakse munamass, tuleks see hoolikalt kõrvaldada.
Egg ropes are collected from the crystallising dish in the breeding cage using tweezers or a blunt pipette.
See tuleks üle kanda väiksele tassile, mis sisaldab paljundusvee proovi.
Each egg rope is placed into a vessel containing culture medium from the crystallising dish it was collected from (e.g. a well of a 12-well micro-plate together with at least 2,5 ml of medium).
Munamasse kasutatakse uue kasvunõu kasutusele võtmiseks (näiteks 2–4 munamassi nõu kohta) või mürgisuse katsetes.
The vessels with the egg ropes are covered with a lid to prevent significant evaporation.
Esimese kasvujärgu vastsed peaksid kooruma 2–3 päeva pärast.
Egg ropes are kept for observation for at least six days after they have been produced so that they can be classified as fertile or infertile.
Uute kasvunõude ülesseadmine
For starting the 2nd generation, at least three but preferably six fertile egg ropes are selected from each breeding cage and together with some food allowed to hatch.
Kui kultuurid on loodud, peaks iga nädal (või katsenõuetest olenevalt väiksema sagedusega) olema võimalik vastsete uute kasvunõude ülesseadmine, kõrvaldades vanemad nõud pärast täiskasvanud sääskede väljaarenemist.
These egg ropes should have been produced at the peak of oviposition, which normally occurs around test day 19 in the controls.
Seda süsteemi kasutades saadakse minimaalse vaevaga regulaarne valmikute varu.
Ideally, the 2nd generation of all treatments is initiated on the same day, but due to chemical related effects on larval development, this may not always be possible.
Katselahuste M4 ja M7 valmistamine
In such a case, the higher concentrations may be initiated later than the lower treatments and the (solvent) control.
Elendt (1990) on kirjeldanud kasvukeskkonda M4.
a.
Kasvukeskkond M7 valmistatakse samal viisil kui kasvukeskkond M4, v.a tabelis 1 nimetatud ainete puhul, mille kontsentratsioon kasvukeskkonnas M7 on kasvukeskkonnaga M4 võrreldes neli korda madalam.
In the spiked water design, the sediment-water system for the 2nd generation is prepared by spiking the test chemical into the overlying water column ca.
Katselahust ei tohiks koostada Elendti ja Biasi (1990) järgi, kuna põhilahuste valmistamiseks ei ole antud NaSiO3 · 5H2O, NaNO3, KH2PO4 ja K2HPO4 kontsentratsioon sobiv.
1 hour before adding the first instar larvae to the test vessels.
Kasvukeskkonna M7 valmistamine
Small volumes of the test chemical solutions are applied below the surface of the water using a pipette.
Iga põhilahus (I) valmistatakse eraldi ja kombineeritud põhilahus (II) valmistatakse kõnealustest põhilahustest (I) (vt tabel 1).
After spiking, slight aeration is supplied.
Kasvukeskkonna M7 valmistamiseks võetakse 50 ml kombineeritud põhilahust (II), lisatakse iga makrotoitaine põhilahuse kogused, mis on esitatud tabelis 2, ja täiendatakse deioniseeritud veega 1 liitrini.
b.
Vitamiinide põhilahuse valmistamiseks lisatakse deioniseeritud veele kolme vitamiini, nagu on näidatud tabelis 3, ja lõplikule M7 kasvukeskkonnale lisatakse vahetult enne selle kasutamist 0,1 ml kombineeritud vitamiinide põhilahust.
In the spiked sediment design, the exposure vessels containing the sediment-water system for the 2nd generation are prepared in the same way as for the 1st generation.
Vitamiinide põhilahust säilitatakse külmutatult väikeste alikvootidena.
Twenty first instar larvae (maximum 48 h post hatching) of the 2nd generation are allocated randomly to each test vessel containing the spiked sediment-water system, using a blunt pipette.
Kasvukeskkonda aereeritakse ja see stabiliseeritakse.
Aeration of the water should be stopped while adding the first instar larvae to the test vessels and remain so for another 24 hours after addition of the larvae.
Kasvukeskkondade M4 ja M7 mikroelementide põhilahused
According to the test design used (see paragraphs 19 and 20), the number of larvae used per concentration is at least 120 (6 replicates per concentration) for the ECX approach and 160 for the NOEC approach (8 replicates per concentration).
Põhilahus (I)
It is necessary to feed the larvae in the test vessels, preferably daily or at least three times per week.
Kogus (mg), mis täiendatakse deioniseeritud veega 1 liitrini
Fish-food (a suspension in water or finely ground food, e.g. Tetra-Min or Tetra-Phyll; see details in Appendix 2) of 0,25 - 0,5 mg (0,35 - 0,5 mg for C. yoshimatsui) per larvae per day is an adequate amount of food for young larvae during the first 10 days of their development.
Kombineeritud põhilahuse (II) valmistamiseks:
Slightly more food may be necessary for older larvae:
segage põhilahuste (I) järgmised kogused (ml) ja täiendage need deioniseeritud veega 1 liitrini
0,5 - 1,0 mg per larvae per day should be sufficient for the rest of the test.
Lõppkontsentratsioonid katselahustes (mg/l)
The food ration should be reduced in all treatments and control if fungal growth is seen or if mortality is observed in controls.
MnCl2 · 4H2O [1]
If fungal development cannot be stopped the test should be repeated.
SrCl2 · 6H2O [1]
The toxicological relevance of exposure via ingestion is generally higher in chemicals with a high affinity for organic carbon or chemicals covalently binding to the sediment.
MoO4 · 2H2O [1]
Hence, when testing chemicals with such properties, the amount of food necessary to ensure survival and natural growth of the larvae may be added to the formulated sediment before the stabilisation period, depending on the regulatory demand.
CuCl2 · 2H2O [1]
To prevent deterioration of the water quality, plant material should be used instead of fish food, e.g. addition of 0,5 % (dry weight) finely ground leaves of stinging nettle (Urtica dioica), mulberry (Morus alba), white clover (Trifolium repens), spinach (Spinacia oleracea) or other plant material (Cerophyl or α-cellulose).
CaCl2 · 6H2O
Addition of the complete ration of an organic food source to the sediment before spiking is not trivial with respect to water quality and biological performance (21), nor a standardised method, but recent studies provide indications that this method works (19) (26).
EDTA · 2H2O [1] [2]
Adult midges in the breeding cage need no feeding normally, but fecundity and fertility are enhanced when a cotton wool pad soaked in a saturated sucrose solution is offered as a food source for emerged adults (34).
FeSO4 · 7H2O [1] [2]
Gentle aeration of the overlying water in the test vessels is supplied 24 hours after addition of the first instar larvae of both generations and is continued throughout the test (care should be taken that the dissolved oxygen concentration does not fall below 60 % of ASV).
Kasvukeskkondade M4 ja M7 makrotoitainete põhilahused
Aeration is provided through a glass Pasteur pipette of which the outlet is fixed 2-3 cm above the sediment layer giving a few bubbles/sec.
Kogus, mis täiendatakse deioniseeritud veega 1 liitrini
When testing volatile chemicals, consideration should be given not to aerate the sediment-water system, while at the same time the validity criterion of minimal 60 % ASV (paragraph 10) should be fulfilled.
Kasvukeskkonna M4 ja M7 valmistamiseks lisatav makrotoitainete põhilahuste kogus
Further guidance is provided in (16).
Lõppkontsentratsioonid katselahustes M4 ja M7
The test with C. riparius is conducted at a constant temperature of 20 °C (± 2 °C).
Kasvukeskkondade M4 ja M7 vitamiinide põhilahus
For C. dilutus and C. yoshimatsui, recommended temperatures are 23 °C and 25 °C (± 2 °C), respectively.
Ühe vitamiinide põhilahuse valmistamiseks valatakse kokku kõik kolm vitamiinilahust.
A 16 hours photoperiod is used and the light intensity should be 500 to 1000 lux.
Kasvukeskkonna M4 ja M7 valmistamiseks lisatav vitamiinide põhilahuse kogus
For the breeding cages an additional one hour dawn and dusk phase may be included.
Tiamiinvesinikkloriid
Exposure duration
Tsüanokobalamiin (B12)
Spiked water design:
Biotiin
The exposure period of the 1st generation starts when the test chemical is spiked into the overlying water of the test vessels (which is one day after insertion of the larvae — for possible amendments of the exposure design, see paragraph 7).
Need ained on M4 ja M7 puhul erinevad, nagu on eespool viidatud.
Exposure of the 2nd larval generation starts immediately, since they are inserted into a sediment-water system that has been already spiked.
Need lahused valmistatakse eraldi, valatakse seejärel kokku ja autoklaavitakse viivitamata.
The maximum exposure duration for the 1st generation is 27 days and for the 2nd generation 28 days (the 1st generation larvae spend one day in the vessels without exposure) for C. riparius and C. yoshimatsui.
Spetsiaalselt koostatud sette valmistamine
Considering the overlap, the complete test duration is approximately 44 days.
SETTE KOOSTIS
For C. dilutus, maximum exposure durations are 64 and 65 days, for the 1st and 2nd generation, respectively.
Spetsiaalselt koostatud sette koostis peaks olema järgmine:
The total duration is approximately 100 days.
Turbasamblaturvas, mille pH on võimalikult lähedal vahemikule 5,5–6,0, nähtavad taimeosad puuduvad, peeneks jahvatatud (osakese suurus ≤ 1 mm) ja õhu käes kuivatatud
Spiked sediment design:
4–5
exposure starts with the addition of the larvae and is maximum 28 days for both generations for C. riparius and C. yoshimatsui and maximum 65 days for both generations for C. dilutus.
> 50 % osakestest peaksid olema vahemikus 50–200 μm
Emergence
CaCO3, pulbriline, keemiliselt puhas
Development time and the total number of fully emerged and alive male and female midges are determined for both generations.
0,05–0,1
Males are easily identified by their plumose antennae and thin body posture.
Juhtivus ≤ 10 μS/cm
Test vessels of both generations should be observed at least three times per week to make visual assessment of any abnormal behaviour of the larvae (e.g. leaving sediment, unusual swimming), compared to the control.
VALMISTAMINE
During the period of emergence, which starts about 12 days after insertion of the larvae for C. riparius and C. yoshimatsui (after 20 days for C. dilutus), emerged midges are counted and sexed at least once, but preferably twice a day (early morning and late afternoon).
Suspensiooni konditsioneeritakse vähemalt kaks päeva temperatuuril 20 ± 2 °C kerge segamisega, et stabiliseerida pH ja luua stabiilne mikroobikomponent. pH mõõdetakse uuesti ja see peaks olema 6,0 ± 0,5.
After identification, the midges of the 1st generation are carefully removed from the vessels and transferred to a breeding cage.
Lõpliku segu pH mõõdetakse uuesti ja reguleeritakse vajaduse korral CaCO3 abil vahemikku 6,5–7,5.
Midges of the 2nd generation are removed and killed after identification.
Enne, kui spetsiaalselt koostatud setet kasutatakse surusääsklastele mürgisuse katses, on soovitatav seda seitsme päeva jooksul konditsioneerida samades tingimustes, mis valdavalt esinevad edasise katse käigus.
Any egg ropes deposited in the test vessels of the 1st generation should be collected individually and transferred with at least 2,5 ml native water to 12-well microplates (or other suitable vessels) which are covered with a lid to prevent significant evaporation.
SÄILITAMINE
The number of dead larvae and visible pupae that have failed to emerge should also be recorded.
Tehisliku sette valmistamiseks kasutatavaid kuivi koostisosi võib säilitada kuivas ja jahedas kohas toatemperatuuril.
Examples of a breeding cage, test vessel and exhauster are provided in Appendix 5.
Spetsiaalselt koostatud (märga) setet ei tohiks enne selle katses kasutamist säilitada.
Reproduction
Seda tuleks kasutada viivitamata pärast seitsmepäevast kohandamisperioodi, millega selle valmistamine lõpetatakse.
Effects on reproduction are assessed via the number of egg ropes produced by the 1st generation of midges and the fertility of these egg ropes.
Kasutuskõlbliku lahjendusvee keemilised omadused
Once per day the egg ropes are collected from the crystallising dish that is placed in each breeding container.
KOOSTISOSA
The egg ropes should be collected and transferred with at least 2,5 ml native water to a 12-wells microplate (one egg rope in each well) or other suitable vessels, which are covered with a lid to prevent significant evaporation.
KONTSENTRATSIOONID
The following characteristics are documented for each egg rope:
Karedus CaCO3-na
day of production, size (normal, i.e. 1,0 ± 0,3 cm or small; typically ≤ 0,5 cm), and structure (normal = banana-form with spiralled egg string or abnormal, e.g. unspiralled egg string) and fertility (fertile or infertile).
Kui kahtlustatakse karedust tekitavate ioonide ja uuritava aine omavahelist mõju, tuleks kasutada väiksema karedusega vett (ja seega ei tohiks nimetatud olukorras kasutada Elendti kasvukeskkonda M4).
Over the course of six days after it was produced the fertility of an egg rope is assessed.
Juhised katse tegemiseks