natgillin/translations-raw huggingface dataset:
If static renewal methods are used, then the stability of the test chemical concentration should be documented and remain within the performance criteria limits.
Ak sa zistí nenormalita alebo heterogenita rozptylu, je potrebné vykonať normalizačnú transformáciu na stabilizáciu rozptylu.
Twenty-four hour renewal periods are recommended.
Ak majú údaje (možno po transformácii) normálnu distribúciu s homogénnym rozptylom, významný vplyv chemikálie sa stanoví pomocou Dunnettovho testu.
Renewal periods exceeding 72 hours are not acceptable.
Ak majú údaje (možno po transformácii) normálnu distribúciu s heterogénnym rozptylom, významný vplyv chemikálie sa stanoví pomocou Tamhaneho-Dunnettovho alebo T3 testu alebo pomocou Mann-Whitneyho-Wilcoxonovho U testu.
Additionally, water quality parameters (e.g. DO, temperature, pH, etc.) should be measured at the end of each renewal period, immediately prior to renewal.
Významná úmrtnosť sa neočakáva, ale treba ju vyhodnotiť pomocou zostupného Cochranovho-Armitageovho testu ak údaje zodpovedajú monotónnosti odozva na dávku a v ostatných prípadoch pomocou Fisherovho exaktného testu s Bonferroniho-Holmovou úpravou.
Test Concentration Range
Významný vplyv chemikálie na vývinové štádium sa stanovuje pomocou zostupnej aplikácie Jonckheereho-Terpstrovho testu na mediány replikátov.
There is a required minimum of three test concentrations and a clean water control (and vehicle control if necessary).
Na stanovenie vplyvu sa odporúča použiť aj multikvantový Jonckheereho test od 20. do 80. percentilu, lebo zohľadňuje zmeny v distribučnom profile.
The minimum test concentration differential between the highest and lowest should be about one order of magnitude.
Vhodnou jednotkou analýzy je replikát, aby údaje predstavovali mediány pre replikáty ak sa používa Jonckheereho-Terpstrov alebo Mann-Whitneyho U test alebo priemery pre replikáty ak sa používa Dunnettov test.
The maximum dose separation is 0,1 and the minimum is 0,33.
Štatistická významnosť všetkých uvedených testov sa posudzuje na hladine významnosti 0,05.
Test Initiation and Conduct
Obrázok 4 predstavuje vývojový diagram vykonávania štatistických testov na spojitých údajoch.
The exposure should be initiated when a sufficient number of tadpoles in the pre-exposure stock population have reached developmental stage 51, according to Nieuwkoop and Faber (8), and which are less than or equal to 17 days of age post fertilisation.
Vývojový diagram štatistických prístupov v prípade údajov o spojitej odozve
For selection of test animals, healthy and normal looking tadpoles of the stock population should be pooled in a single vessel containing an appropriate volume of dilution water.
Osobitné aspekty analýzy údajov
For developmental stage determination, tadpoles should be individually removed from the pooling tank using a small net or strainer and transferred to a transparent measurement chamber (e.g. 100 mm Petri dish) containing dilution water.
Používanie úrovní narušenia skupiny s určitou koncentráciou chemikálie
For stage determination, it is preferred not to use anaesthesia, however one may individually anaesthetise the tadpoles using 100 mg/l tricaine methanesulfonate (e.g. MS-222), appropriately buffered with sodium bicarbonate (pH 7,0), before handling.
Pri rozhodovaní, či replikát alebo celá skupina s určitou koncentráciou chemikálie vykazuje zjavnú toxicitu a mala by sa vylúčiť z analýzy, sa zohľadňuje viacero faktorov.
If used, methodology for appropriately using e.g. MS-222 for anaesthesia should be obtained from experienced laboratories and reported with the test results.
Zjavná toxicita je vymedzená > 2 úhynmi v ktoromkoľvek z replikátov, ktoré možno vysvetliť iba toxicitou a nie technickou chybou.
Animals should be carefully handled during this transfer in order to minimise handling stress and to avoid any injury.
Iné známky zjavnej toxicity zahŕňajú krvácanie abnormálne správanie abnormálne vzory plávania anorexiu a akékoľvek iné klinické známky choroby.
The developmental stage of the animals is determined using a binocular dissection microscope.
V prípade subletálnych známok toxicity môžu byť potrebné kvalitatívne hodnotenia, ktoré by sa vždy mali vykonať porovnaním s kontrolnou skupinou s čistou vodou.
To reduce the ultimate variability in developmental stage, it is important that this staging be conducted as accurately as possible.
Kontrolné skupiny s rozpúšťadlom
According to Nieuwkoop and Faber (8), the primary developmental landmark for selecting stage 51 organisms is hind limb morphology.
O použití rozpúšťadla by sa malo uvažovať až ako o poslednej možnosti ak sa zvážili všetky ostatné možnosti aplikovania chemikálie.
The morphological characteristics of the hind limbs should be examined under the microscope.
Ak sa použije rozpúšťadlo, mala by sa zároveň vykonať analýza kontrolnej skupiny s čistou vodou.
While the complete Nieuwkoop and Faber (8) guide should be consulted for comprehensive information on staging tadpoles, one can reliably determine stage using prominent morphological landmarks.
Po skončení testu by sa malo vykonať hodnotenie potenciálneho vplyvu rozpúšťadla.
The following table can be used to simplify and standardise the staging process throughout the study by identifying those prominent morphological landmarks associated with different stages, assuming that development is normal.
To sa dosiahne štatistickým porovnaním kontrolnej skupiny s rozpúšťadlom a kontrolnej skupiny s čistou vodou.
Prominent morphological staging landmarks based on Neuwkoop and Faber guidance
Najdôležitejšie sledované parametre, ktoré treba zohľadniť v tejto analýze, sú vývinové štádium, SVL a mokrá hmotnosť, lebo môžu byť ovplyvnené netyreoideálnou toxicitou.
Prominent Morphological Landmarks
Ak sa zistia štatisticky významné rozdiely v týchto sledovaných parametroch medzi kontrolnou skupinou s čistou vodou a kontrolnou skupinou s rozpúšťadlom, stanovte sledované parametre štúdie pre merania odozvy s použitím kontrolnej skupiny s čistou vodou.
Developmental Stage
Ak sa nezistí štatisticky významný rozdiel medzi kontrolnou skupinou s čistou vodou a kontrolnou skupinou s rozpúšťadlom v žiadnej z nameraných závisle premenných, stanovte sledované parametre štúdie pre merania odozvy s použitím spojenej kontrolnej skupiny s riediacou vodou a rozpúšťadlom.
Hindlimb
Skupiny s aplikovanou chemikáliou, ktoré dosiahli vývinové štádium 60 a vyššie
Forelimb
Jedným prístupom je použiť na štatistické analýzy mokrej hmotnosti a/alebo SVL len žubrienky s vývinovým štádiom 60 alebo nižším.
Craniofacial structure
Predpokladá sa, že tento prístup poskytuje dostatočne spoľahlivé informácie o závažnosti možných vplyvov na rast, pokiaľ sa z analýz odstráni iba malá časť testovacích živočíchov (≤ 20 %).
Olfactory nerve morphology
Ak sa vývin po štádiu 60 zistí pri zvýšenom počte žubrienok (≥ 20 %) v prípade jednej alebo viacerých nominálnych koncentrácií, mala by sa na všetkých žubrienkach vykonať dvojfaktorová ANOVA s hierarchickou štruktúrou rozptylu na vyhodnotenie vplyvu na rast v dôsledku aplikovania chemikálie so zohľadnením vplyvu vývinu po neskoré štádium na rast.
Tail length
Dodatok 3 obsahuje usmernenie o dvojfaktorovej analýze ANOVA hmotnosti a dĺžky.
For test initiation, all tadpoles should be at stage 51.
Series on Testing and Assessment.
The most prominent morphological staging landmark for that stage is hind limb morphology, which is demonstrated in Figure 1.
No.
Hind limb morphology of a stage 51 X. laevis tadpole
77, Paríž.
In addition to the developmental stage selection, an optional size selection of the experimental animals may be used.
E 1439-98.
For this purpose, the whole body length (not SVL) should be measured at day 0 for a sub-sample of approximately 20 NF stage 51 tadpoles.
(ed.) academic Press, New York, s.
After calculation of the mean whole body length for this group of animals, minimum and maximum limits for the whole body length of experimental animals can be set by allowing a range of the mean value ± 3 mm (mean values of whole body length range between 24,0 and 28,1 mm for stage 51 tadpoles).
467 – 599.
However, developmental staging is the primary parameter in determining the readiness of each test animal.
Podmienky pokusu pre 21-dňovú skúšku metamorfózy obojživelníkov
Tadpoles exhibiting grossly visible malformations or injuries should be excluded from the assay.
Testovací živočích
Tadpoles that meet the stage criteria described above are held in a tank of clean culture water until the staging process is completed.
Larvy Xenopus laevis
Once the staging is completed, the larvae are randomly distributed to exposure treatment tanks until each tank contains 20 larvae.
Počiatočné larválne štádium
Each treatment tank is then inspected for animals with abnormal appearance (e.g., injuries, abnormal swimming behaviour, etc.).
Štádium 51 podľa Nieuwkoopa a Fabera
Overtly unhealthy looking tadpoles should be removed from the treatment tanks and replaced with larvae newly selected from the pooling tank.
Obdobie expozície
For more in-depth information on test termination procedures and processing of tadpoles, refer to the OECD Guidance Document on Amphibian Thyroid Histology (9).
21 dní
The random procedure used should give each organism on test equal probability of being selected.
Testované koncentrácie
Tadpoles not selected are returned to the tank of origin and the selected tadpoles are humanely euthanised in 150 to 200 mg/l e.g. MS-222, appropriately buffered with sodium bicarbonate to achieve pH 7,0.
Režim expozície
The euthanised tadpoles are rinsed in water and blotted dry, followed by body weight determination to the nearest milligram.
Prietokový (prednostne) a/alebo statický obnovovací
Hind limb length, snout to vent length, and developmental stage (using a binocular dissection microscope) are determined for each tadpole.
Prietok testovacieho systému
Day 21 Measurements (Test Termination)
25 ml/min. (výmena celého objemu približne každých 2,7 hod.)
At test termination (day 21), the remaining tadpoles are removed from the test tanks and humanely euthanised in 150 to 200 mg/l e.g. MS-222, appropriately buffered with sodium bicarbonate, as above.
Základné sledované parametre/dni stanovenia
Tadpoles are rinsed in water and blotted dry, followed by body weight determination to the nearest milligram.
a 21. deň
Developmental stage, SVL, and hind limb lengths are measured for each tadpole.
Histológia štítnej žľazy
All larvae are placed in Davidson's fixative for 48 to 72 hours either as whole body samples or as trimmed head tissue samples containing the lower jaw for histological assessments.
Riediaca voda/laboratórna kontrola
For histopathology, a total of five tadpoles should be sampled from each replicate tank.
Odchlórovaná voda z vodovodu (filtrovaná pomocou drevného uhlia) alebo rovnocenný laboratórny zdroj
Since follicular cell height is stage dependent (10), the most appropriate sampling approach for histological analyses is to use stage-matched individuals, whenever possible.
Hustota lariev
In order to select stage-matched individuals, all larvae should first be staged prior to selection and subsequent processing for data collection and preservation.
20 lariev/testovaciu nádobu (5/l)
This is necessary because normal divergence in development will result in differential stage distributions within each replicate tank.
Testovací roztok/testovacia nádoba
Animals selected for histopathology (n = 5 from each replicate) should be matched to the median stage of the controls (pooled replicates) whenever possible.
4 – 10 l (minimálna hĺbka vody 10 – 15 cm)/testovacia nádoba zo skla alebo nehrdzavejúcej ocele (napr. 22,5 cm × 14 cm × 16,5 cm)
If there are replicate tanks with more than five larvae at the appropriate stage, then five larvae are randomly selected.
Opakovanie
That is, if the chemical treatment is associated with a retardation of development, then additional larvae should be sampled from the next lower stage.
≤ 10 % na testovaciu nádobu s replikátom
In turn, if the chemical treatment is associated with an acceleration of development, then additional larvae should be sampled from the next upper stage.
Fixácia štítnej žľazy
In cases of severe alterations of tadpole development due to treatment with a test chemical, there might be no overlap of the stage distribution in the chemical treatments with the calculated control median developmental stage.
Počet fixovaných jedincov
In only these cases, the selection process should be modified by using a stage different from the control median stage to achieve a stage-matched sampling of larvae for thyroid histopathology.
Všetky žubrienky (5/replikát sa vyhodnotí na začiatku)
Furthermore, if stages are indeterminate (i.e., asynchrony), then 5 tadpoles from each replicate should be randomly chosen for histological analysis.
Oblasť
The rationale underlying sampling of any larvae that are not at a stage equivalent to the control median developmental stage should be reported.
Hlava alebo celé telo
Determination of Biological Endpoints
Fixačná kvapalina
During the 21 day exposure phase, measurement of primary endpoints is performed on days 7 and 21, however daily observation of test animals is necessary.
Davidsonov fixačný prostriedok
Table 3 provides an overview of the measurement endpoints and the corresponding observation time points.
Potrava
More detailed information for technical procedures for measurement of apical endpoints and histological assessments is available in the OECD guidance documents (9).
Sera Micron® alebo rovnocenná
Observation time points for primary endpoints in the AMA
Množstvo/frekvencia
Apical Endpoints
Kŕmny režim pre potravu Sera Micron® sa nachádzajú v tabuľke 1.
Daily
Osvetlenie
Mortality
Fotoperióda
Hind Limb Length
12 hod. svetlo:
Snout-Vent Length
12 hod. tma
Wet Body Weight
Intenzita
Thyroid Gland Histology
600 – 2000 luxov (merané na povrchu vody)
Developmental stage, hind limb length, SVL and wet weight are the apical endpoints of the AMA, and each is briefly discussed below.
Teplota vody
Further technical information for collecting these data is available in the guidance documents referenced including procedures for computer-assisted analysis which are recommended for use.
6,5 – 8,5
The developmental stage of X. laevis tadpoles is determined using the staging criteria of Nieuwkoop and Faber (8).
Koncentrácia rozpusteného kyslíka
Developmental stage data are used to determine if development is accelerated, asynchronous, delayed or unaffected.
> 3,5 mg/l (> 40 % rozpustnosť vzdušného kyslíka)
Acceleration or delay of development is determined by making a comparison between the median stage achieved by the control and treated groups.
Harmonogram odoberania vzoriek na chemickú analýzu
Asynchronous development is reported when the tissues examined are not malformed or abnormal, but the relative timing of the morphogenesis or development of different tissues is disrupted within a single tadpole.
Raz za týždeň (4 odobratia vzorky/test)
Differentiation and growth of the hind limbs are under control of thyroid hormones and are major developmental landmarks already used in the determination of developmental stage.
Oznamovacie tabuľky pre nespracované údaje a súhrnné údaje
Hind limb development is used qualitatively in the determination of developmental stage, but is considered here as a quantitative endpoint.
Všeobecné informácie o testovanej chemikálii
Therefore, hind limb length is measured as an endpoint to detect effects on the thyroid axis (Figure 2).
Informácie o chemikálii
For consistency, hind limb length is measured on the left hind limb.
Uveďte testovanú chemikáliu, jednotky koncentrácie a testované koncentrácie
Hind limb length is evaluated both at day 7 and at day 21 of the test.
Jednotky koncentrácie:
On day 7, measuring hind limb length is straightforward, as illustrated in Figure 2.
Testovaná koncentrácia 1
However, measuring hind limb length on day 21 is more complicated due to bends in the limb.
Testovaná koncentrácia 2
Therefore, measurements of hind limb length at day 21 should originate at the body wall and follow the midline of the limb through any angular deviations.
Testovaná koncentrácia 3
Changes in hind limb length at day 7, even if not evident at day 21, are still considered significant for potential thyroid activity.
Testovaná koncentrácia 4
Length measurements are acquired from digital photographs using image analysis software as described in the OECD Guidance Document on Amphibian Thyroid Histology (9).
Dátum (0. deň):
Body Length and Wet Weight
Uveďte dátum (mm/dd/rr)
Determinations of snout to vent length (SVL) (Figure 2) and wet weight are included in the test protocol to assess possible effects of test chemicals on the growth rate of tadpoles in comparison to the control group and are useful in detecting generalised toxicity to the test chemical.
Dátum (7. deň):
Because the removal of adherent water for weight determinations can cause stressful conditions for tadpoles and may cause skin damage, these measurements are performed on the day 7 sub-sampled tadpoles and all remaining tadpoles at test termination (day 21).
Dátum (21. deň):
For consistency, use the cranial aspect of the vent as the caudal limit of the measurement.
Hárky na zaznamenanie nespracovaných údajov v 7. a 21. deň
Snout to vent length (SVL) is used to assess tadpole growth as illustrated in Figure 2.
DEŇ X
Types of body length measurements and (B) Hind limb length measurements for X. laevis tadpoles (1)
DÁTUM 00/00/00
While developmental stage and hind limb length are important endpoints to evaluate exposure-related changes in metamorphic development, developmental delay cannot, by itself, be considered a diagnostic indicator of anti-thyroidal activity.
Číslo koncentrácie
Some changes may only be observable by routine histopathological analysis.
Číslo replikátu
Diagnostic criteria include thyroid gland hypertrophy/atrophy, follicular cell hypertrophy, follicular cell hyperplasia, and as additional qualitative criteria:
Číslo jedinca
follicular lumen area, colloid quality and follicular cell height/shape.
Identifikátor jedinca
Severity grading (4 grades) should be reported.
Dĺžka SVL (mm)
Information on obtaining and processing samples for histological analysis and for performing histologic analyses on tissue samples is available in “Amphibian Metamorphosis Assay: Part 1 — Technical guidance for morphologic sampling and histological preparation” and “Amphibian Metamorphosis Assay: Part 2 — Approach to reading studies, diagnostic criteria, severity grading and atlas” (9).
Dĺžka zadných končatín (mm)
Laboratories performing the assay for the first time(s) should seek advice from experienced pathologists for training purpose prior to undertaking histological analysis and evaluation of the thyroid gland.
Mokrá hmotnosť celého organizmu (mg)
Overt and significant changes in apical endpoints indicating developmental acceleration or asynchrony may preclude the necessity to perform histopathological analysis of the thyroid glands.
Vypočítané zhrnutia údajov o sledovaných parametroch zo 7. a 21. dňa
However, absence of overt morphological changes or evidence of developmental delay warrants histological analyses.
Výpočty hodnôt v bunkách sú prepojené s údajmi v tabuľke 2.
All test tanks should be checked daily for dead tadpoles and the numbers recorded for each tank.
Hmotnosť (mg) TRT REP
The date, concentration and tank number for any observation of mortality should be recorded.
MIN.
Dead animals should be removed from the test tank as soon as observed.
MEDIÁN
Mortality rates exceeding 10 % may indicate inappropriate test conditions or toxic effects of the test chemical.
MAX.
Additional Observations
PRIEMER
Cases of abnormal behaviour and grossly visible malformations and lesions should be recorded.
ŠTD.
The date, concentration and tank number for any observation of abnormal behaviour, gross malformations or lesions should be recorded.
ŠTD.
Normal behaviour is characterised by the tadpoles being suspended in the water column with tail elevated above the head, regular rhythmic tail fin beating, periodic surfacing, operculating, and being responsive to stimulus.
ODCH.
Abnormal behaviour would include, for example, floating on the surface, lying on the bottom of the tank, inverted or irregular swimming, lack of surfacing activity, and being nonresponsive to stimulus.
Údaje o dennej úmrtnosti
In addition, gross differences in food consumption between treatments should be recorded.
Výpočty hodnôt v bunkách sú prepojené s údajmi v tabuľke 1.
Gross malformations and lesions could include morphological abnormalities (e.g. limb deformities), hemorrhagic lesions, bacterial or fungal infections, to name a few.
Deň testu
These determinations are qualitative and should be considered akin to clinical signs of disease/stress and made in comparison to control animals.
Počet replikátov
If the occurrence or rate of occurrence is greater in exposed tanks than in the controls, then these should be considered as evidence for overt toxicity.
Počet aplikovaní chemikálie
Data Collection
Tabuľka 5
All data should be collected using electronic or manual systems which conform to good laboratory practices (GLP).
Kritériá kvality vody
Characterisation of the test chemical:
Expozičný systém (prietokový/statický obnovovací):
physical-chemical properties;
Intenzita svetla:
information on stability and biodegradability;
Cyklus svetla a tmy:
Chemical information and data:
Potrava:
method and frequency of preparation of dilutions.
Dávka krmiva:
Test chemical information includes actual and nominal concentrations of the test chemical, and in some cases, non-parent chemical, as appropriate.
pH vody:
Test chemical measurements may be required for stock solutions as well as for test solutions;
Koncentrácia jódu v testovacej vode:
Solvent (if other than water):
Tabuľka 6
justification of the choice of solvent, and characterisation of solvent (nature, concentration used);
Súhrnné chemické údaje
Operational records:
Názov chemikálie:
these consist of observations pertaining to the functioning of the test system and the supporting environment and infrastructure.
Č. CAS:
Typical records include:
Tabuľka 7
ambient temperature, test temperature, photoperiod, status of critical components of the exposure system (e.g. pumps, cycle counters, pressures), flow rates, water levels, stock bottle changes, and feeding records.
Histopatologické oznamovacie tabuľky pre hlavné kritériá
General water quality parameters include:
Dátum:
pH, DO, conductivity, total iodine, alkalinity, and hardness;
Chemikália:
Deviations from the test method:
Patológ:
this information should include any information or narrative descriptions of deviations from the test method;
Hypertrofia štítnej žľazy
Biological observations and data:
Atrofia štítnej žľazy
these include daily observations of mortality, food consumption, abnormal swimming behaviour, lethargy, loss of equilibrium, malformations, lesions, etc.
Hypertrofia folikulárnych buniek
Observations and data collected at predetermined intervals include:
Hyperplázia folikulárnych buniek
developmental stage, hind limb length, snout vent length, and wet weight;
ID kontrolného živočícha – replikát 1
Statistical analytical techniques and justification of techniques used;
ID živočícha dostávajúceho dávku – replikát 1
results of the statistical analysis preferably in tabular form;
ID kontrolného živočícha – replikát 2
Histological data:
ID živočícha dostávajúceho dávku – replikát 2
these include narrative descriptions, as well as graded severity and incidence scores of specific observations, as detailed in the histopathology guidance document;
Spolu:
Ad hoc observations:
Tabuľka 8
these observations should include narrative descriptions of the study that do not fit into the previously described categories.
Ďalšie histopatologické kritériá
Data reporting
Zväčšenie plochy folikulárnej dutiny
Appendix 2 contains daily data collection spreadsheets that can be used as guidance for raw data entry and for calculations of summary statistics.
Zmenšenie plochy folikulárnej dutiny
Additionally, reporting tables are provided that are convenient for communicating summaries of endpoint data.
Tabuľka 9