natgillin/translations-raw huggingface dataset:
Jsou-li údaje (případně po transformaci) rozděleny normálně s heterogenním rozptylem, stanoví se významný účinek expozice Tamhaneovým-Dunnettovým testem nebo testem T3 nebo Mannovým-Whitneyovým-Wilcoxonovým U-testem.
Once the staging is completed, the larvae are randomly distributed to exposure treatment tanks until each tank contains 20 larvae.
Významná mortalita se nepředpokládá, ale měla by být posouzena pomocí Cochranova-Armitageova testu sestupného trendu, pokud údaje odpovídají monotónní závislosti dávka-odezva, a pokud neodpovídají, pomocí Fisherova exaktního testu s Bonferroni-Holmovou korekcí.
Each treatment tank is then inspected for animals with abnormal appearance (e.g., injuries, abnormal swimming behaviour, etc.).
Významný účinek expozice pro stadium vývoje se stanoví pomocí Jonckheerova-Terpstrova testu sestupného trendu aplikovaného na mediánové hodnoty opakování.
Overtly unhealthy looking tadpoles should be removed from the treatment tanks and replaced with larvae newly selected from the pooling tank.
Alternativně a přednostně by pro stanovení účinku od 20. do 80. percentilu měl být použit Jonckheerův multikvantový test, jelikož zohledňuje změny v profilu distribuce.
For more in-depth information on test termination procedures and processing of tadpoles, refer to the OECD Guidance Document on Amphibian Thyroid Histology (9).
Při méně než pěti opakováních při každé expozici nebo kontrole by měly být použity exaktní permutační verze Jonckheerova-Terpstrova testu a Mannova-Whitneyova testu, jsou-li dostupné.
The random procedure used should give each organism on test equal probability of being selected.
Na obrázku 4 je vývojový diagram pro provádění statistických testů kontinuálních údajů.
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.
Vývojový diagram pro statistické metody zpracování údajů o kontinuální odezvě.
The euthanised tadpoles are rinsed in water and blotted dry, followed by body weight determination to the nearest milligram.
Úvahy o analýze zvláštních dat
Hind limb length, snout to vent length, and developmental stage (using a binocular dissection microscope) are determined for each tadpole.
Použití zpochybněných úrovní expozice
Day 21 Measurements (Test Termination)
Při určování toho, zda opakování nebo celá expozice vykazují zjevnou toxicitu a měly by být z analýzy vyřazeny, se zvažuje několik faktorů.
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.
Zjevná toxicita je definována jako > 2 úhyny v každém opakování, které lze vysvětlit pouze toxicitou, a nikoli technickou chybou.
Tadpoles are rinsed in water and blotted dry, followed by body weight determination to the nearest milligram.
Jiné příznaky zjevné toxicity zahrnují krvácení, nestandardní chování, nestandardní plavání, anorexii a jakékoli jiné klinické příznaky choroby.
Developmental stage, SVL, and hind limb lengths are measured for each tadpole.
Při subletálních příznacích toxicity mohou být nezbytná kvalitativní hodnocení, která by měla být vždy prováděna ve srovnání s kontrolní skupinou v čisté vodě.
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.
Kontroly s rozpouštědlem
For histopathology, a total of five tadpoles should be sampled from each replicate tank.
Použití rozpouštědla by mělo být zvažováno pouze jako poslední možnost, když byly uváženy všechny ostatní možnosti dávkování chemické látky.
Since follicular cell height is stage dependent (10), the most appropriate sampling approach for histological analyses is to use stage-matched individuals, whenever possible.
Je-li použito rozpouštědlo, měla by být souběžně provedena u kontrolní skupiny zkouška s čistou vodou.
In order to select stage-matched individuals, all larvae should first be staged prior to selection and subsequent processing for data collection and preservation.
Při ukončení zkoušky je třeba provést hodnocení případných účinků rozpouštědla.
This is necessary because normal divergence in development will result in differential stage distributions within each replicate tank.
To se provede pomocí statistického srovnání kontrolní skupiny s rozpouštědlem a kontrolní skupiny s čistou vodou.
Animals selected for histopathology (n = 5 from each replicate) should be matched to the median stage of the controls (pooled replicates) whenever possible.
Nejdůležitějšími ukazateli, které je třeba při této analýze posoudit, jsou stadium vývoje, délka těla od tlamy po kloaku a hmotnost ve vlhkém stavu, jelikož tyto ukazatele mohou být ovlivněny netyreoidální toxicitou.
If there are replicate tanks with more than five larvae at the appropriate stage, then five larvae are randomly selected.
Jsou-li zaznamenány statisticky významné rozdíly v těchto ukazatelích mezi kontrolní skupinou s čistou vodou a kontrolní skupinou s rozpouštědlem, stanovte ukazatele studie pro měření odezev pomocí kontrolní skupiny s čistou vodou.
If there are replicate tanks with less than five larvae at the appropriate stage, then randomly selected individuals from the next lower or upper developmental stage should be sampled to reach a total sample size of five larvae per replicate.
Pokud neexistují žádné statisticky významné rozdíly mezi kontrolní skupinou s čistou vodou a kontrolní skupinou s rozpouštědlem pro všechny měřené proměnné odezvy, stanovte ukazatele studie pro měření odezev pomocí sloučené kontrolní skupiny s ředicí vodou a kontrolní skupiny s rozpouštědlem.
Preferably, the decision to sample additional larvae from either the next lower or upper developmental stage should be made based on an overall evaluation of the stage distribution in the control and chemical treatments.
Exponované skupiny dosahující vývojového stadia 60 a vyšších
That is, if the chemical treatment is associated with a retardation of development, then additional larvae should be sampled from the next lower stage.
Jedna metoda spočívá v tom, že pro statistické analýzy hmotnosti ve vlhkém stavu a/nebo délky těla od tlamy po kloaku se posoudí pouze pulci ve stadiu vývoje, které je nižší nebo rovno stadiu 60.
In turn, if the chemical treatment is associated with an acceleration of development, then additional larvae should be sampled from the next upper stage.
Má se za to, že tato metoda poskytuje dostatečně průkazné údaje o závažnosti možných růstových účinků, pokud se z analýzy vyloučí pouze malá část (≤ 20 %) pokusných organismů.
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.
Pokud stadium vývoje vyšší než stadium 60 vykazuje vyšší počet pulců (≥ 20 %) při jedné nebo více nominálních koncentracích, pak by měla být u všech pulců provedena dvoufaktorová hierarchická analýza rozptylu (ANOVA), což umožní posoudit růstové účinky vyvolané expozicí chemické látce při zohlednění účinku pozdního stadia vývoje na růst.
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.
Pokyny týkající se dvoufaktorové analýzy ANOVA hmotnosti a délky obsahuje dodatek 3.
Furthermore, if stages are indeterminate (i.e., asynchrony), then 5 tadpoles from each replicate should be randomly chosen for histological analysis.
Series on Testing and Assessment.
The rationale underlying sampling of any larvae that are not at a stage equivalent to the control median developmental stage should be reported.
No.
Determination of Biological Endpoints
77, Paris.
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.
E 1439-98.
Table 3 provides an overview of the measurement endpoints and the corresponding observation time points.
467–599.
More detailed information for technical procedures for measurement of apical endpoints and histological assessments is available in the OECD guidance documents (9).
Experimentální podmínky pro 21denní zkoušku metamorfózy obojživelníků
Observation time points for primary endpoints in the AMA
Pokusné zvíře
Apical Endpoints
Larvy Xenopus laevis
Daily
Výchozí stadium larev
Mortality
Stadium 51 podle Nieuwkoopa a Fabera
Hind Limb Length
Doba expozice
Snout-Vent Length
21 dnů
Wet Body Weight
Kritéria pro výběr larev
Thyroid Gland Histology
Stadium vývoje a celková délka (volitelné)
Developmental stage, hind limb length, SVL and wet weight are the apical endpoints of the AMA, and each is briefly discussed below.
Zkušební koncentrace
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.
Minimálně 3 koncentrace přibližně v rozmezí jednoho řádu
The developmental stage of X. laevis tadpoles is determined using the staging criteria of Nieuwkoop and Faber (8).
Režim expozice
Developmental stage data are used to determine if development is accelerated, asynchronous, delayed or unaffected.
Průtokový (upřednostňuje se) a/nebo statický s obnovováním média
Acceleration or delay of development is determined by making a comparison between the median stage achieved by the control and treated groups.
Průtok zkušebního systému
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.
25 ml/min (výměna celého objemu přibližně každých 2,7 h)
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.
Hlavní ukazatele / dny určování
Hind limb development is used qualitatively in the determination of developmental stage, but is considered here as a quantitative endpoint.
D 7 a 21
Therefore, hind limb length is measured as an endpoint to detect effects on the thyroid axis (Figure 2).
Histologie štítné žlázy
For consistency, hind limb length is measured on the left hind limb.
Ředicí voda / laboratorní kontrola
Hind limb length is evaluated both at day 7 and at day 21 of the test.
Dechlorovaná vodovodní voda (filtrovaná přes aktivní uhlí) nebo rovnocenný laboratorní zdroj
On day 7, measuring hind limb length is straightforward, as illustrated in Figure 2.
Hustota larev
However, measuring hind limb length on day 21 is more complicated due to bends in the limb.
20 larev / zkušební nádoba (5 / l)
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.
Zkušební roztok / zkušební nádoba
Changes in hind limb length at day 7, even if not evident at day 21, are still considered significant for potential thyroid activity.
4–10 l (minimální obsah vody 10–15 cm) / zkušební nádoba skleněná nebo z nerezové oceli (např. 22,5 cm × 14 cm × 16,5 cm)
Length measurements are acquired from digital photographs using image analysis software as described in the OECD Guidance Document on Amphibian Thyroid Histology (9).
Opakování
Body Length and Wet Weight
4 zkušební nádoby k opakování / zkušební koncentrace a kontrola
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.
Přijatelná mortalita v kontrolních skupinách
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).
≤ 10 % v každé zkušební nádobě k opakování
For consistency, use the cranial aspect of the vent as the caudal limit of the measurement.
Fixace štítné žlázy
Snout to vent length (SVL) is used to assess tadpole growth as illustrated in Figure 2.
Počet fixovaných
Types of body length measurements and (B) Hind limb length measurements for X. laevis tadpoles (1)
Všichni pulci (zpočátku se hodnotí 5/opakování)
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.
Oblast
Some changes may only be observable by routine histopathological analysis.
Hlava nebo celé tělo
Diagnostic criteria include thyroid gland hypertrophy/atrophy, follicular cell hypertrophy, follicular cell hyperplasia, and as additional qualitative criteria:
Fixační kapalina
follicular lumen area, colloid quality and follicular cell height/shape.
Davidsonovo fixační médium
Severity grading (4 grades) should be reported.
Potrava
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).
Sera Micron® nebo rovnocenné
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.
Množství/frekvence
Overt and significant changes in apical endpoints indicating developmental acceleration or asynchrony may preclude the necessity to perform histopathological analysis of the thyroid glands.
Režim krmení při použití Sera Micron® viz tabulka 1
However, absence of overt morphological changes or evidence of developmental delay warrants histological analyses.
Osvětlení
All test tanks should be checked daily for dead tadpoles and the numbers recorded for each tank.
Fotoperioda
The date, concentration and tank number for any observation of mortality should be recorded.
12 h světla:
Dead animals should be removed from the test tank as soon as observed.
12 h tmy
Mortality rates exceeding 10 % may indicate inappropriate test conditions or toxic effects of the test chemical.
Intenzita
Additional Observations
600 až 2000 luxů (měřeno na povrchu vody)
Cases of abnormal behaviour and grossly visible malformations and lesions should be recorded.
Teplota vody
The date, concentration and tank number for any observation of abnormal behaviour, gross malformations or lesions should be recorded.
6,5 – 8,5
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.
Koncentrace rozpuštěného kyslíku (DO)
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.
> 3,5 mg/l (> 40 % nasycení vzduchem)
In addition, gross differences in food consumption between treatments should be recorded.
Rozvrh odběru vzorků pro analytickou chemii
Gross malformations and lesions could include morphological abnormalities (e.g. limb deformities), hemorrhagic lesions, bacterial or fungal infections, to name a few.
Jednou/týden (4 případy odběru vzorků / zkouška)
These determinations are qualitative and should be considered akin to clinical signs of disease/stress and made in comparison to control animals.
Vykazovací tabulka pro naměřené údaje a souhrnné údaje
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.
Chemické informace o hlavní zkoušce
Data Collection
Chemické informace
All data should be collected using electronic or manual systems which conform to good laboratory practices (GLP).
Uveďte zkoušenou chemickou látku, jednotky koncentrace a expozice
Characterisation of the test chemical:
Jednotky koncentrace
physical-chemical properties;
Expozice 1
information on stability and biodegradability;
Expozice 2
Chemical information and data:
Expozice 3
method and frequency of preparation of dilutions.
Expozice 4
Test chemical information includes actual and nominal concentrations of the test chemical, and in some cases, non-parent chemical, as appropriate.
Datum (den 0):
Test chemical measurements may be required for stock solutions as well as for test solutions;
Uveďte datum (mm/dd/rr)
Solvent (if other than water):
Datum (den 7):
justification of the choice of solvent, and characterisation of solvent (nature, concentration used);
Datum (den 21):
Operational records:
Formuláře pro sběr údajů naměřených v den 7 a 21
these consist of observations pertaining to the functioning of the test system and the supporting environment and infrastructure.
DEN X
Typical records include:
DATUM 00/00/00
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.
Číslo expozice
General water quality parameters include:
Číslo opakování
pH, DO, conductivity, total iodine, alkalinity, and hardness;
Číslo jedince
Deviations from the test method:
Identifikátor jedince
this information should include any information or narrative descriptions of deviations from the test method;
Délka těla od tlamy po kloaku (mm)
Biological observations and data:
Délka těla od tlamy po kloaku (mm)
these include daily observations of mortality, food consumption, abnormal swimming behaviour, lethargy, loss of equilibrium, malformations, lesions, etc.
Hmotnost celého organismu ve vlhkém stavu (mg)
Observations and data collected at predetermined intervals include:
Vypočtené souhrnné údaje k ukazatelům ze dne 7 a 21
developmental stage, hind limb length, snout vent length, and wet weight;
Výpočty v buňkách jsou propojeny s údaji zadanými v tabulce 2.
Statistical analytical techniques and justification of techniques used;
Délka zadní končetiny (mm)
results of the statistical analysis preferably in tabular form;
Hmotnost (mg)
Histological data:
EXP
these include narrative descriptions, as well as graded severity and incidence scores of specific observations, as detailed in the histopathology guidance document;
REP
Ad hoc observations:
MIN
these observations should include narrative descriptions of the study that do not fit into the previously described categories.
MEDI
Data reporting
MAX
Appendix 2 contains daily data collection spreadsheets that can be used as guidance for raw data entry and for calculations of summary statistics.
STŘED SD
Additionally, reporting tables are provided that are convenient for communicating summaries of endpoint data.
Údaje o denní mortalitě
Reporting tables for histological assessments can be found in Appendix 2.
Výpočty v buňkách jsou propojeny s údaji zadanými v tabulce 1.
Performance Criteria and Test Acceptability/Validity
Den zkoušky
Generally, gross deviations from the test method will result in unacceptable data for interpretation or reporting.
Počet opakování
Therefore, the following criteria in Table 4 have been developed as guidance for determining the quality of the test performed, the general performance of the control organisms.
Počet expozic
Table 4
Tabulka 5
Performance criteria for the AMA
Kritéria pro kvalitu vody
Criterion
Expoziční systém (průtokový / statický s obnovováním média):
Acceptable limits
Intenzita světla:
Test concentrations
Cyklus světlo-tma:
Maintained at ≤ 20 % CV (variability of measured test concentration) over the 21 day test
Potrava:
Mortality in controls
Množství krmiva:
≤ 10 % — mortality in any one replicate in the controls should not exceed 2 tadpoles
pH vody:
Minimum median developmental stage of controls at end of test
Koncentrace jódu ve zkušební vodě:
Spread of development stage in control group
Tabulka 6
The 10th and the 90th percentile of the development stage distribution should not differ by more than 4 stages
Souhrn údajů o chemickém vyšetření
Dissolved Oxygen
Chemický název
≥ 40 % air saturation [8]
Číslo CAS:
pH should be maintained between 6,5-8,5.
Tabulka 7
The inter-replicate/inter-treatment differentials should not exceed 0,5.
Vykazovací tabulky pro hlavní histopatologická kritéria
± 1 °C — the inter-replicate/inter-treatment differentials should not exceed 0,5 °C
Datum:
Test concentrations without overt toxicity
Chemická látka:
Replicate performance
Patolog:
≤ 2 replicates across the test can be compromised
Hypertrofie štítné žlázy
Special conditions for use of a solvent
Atrofie štítné žlázy
If a carrier solvent is used, both a solvent control and clean water control should be used and results reported
Hyperplazie folikulárních buněk
See below for more information
ID kontrolního zvířete – opakování 1
Special conditions for static renewal system
ID exponovaného zvířete – opakování 1
Representative chemical analyses before and after renewal should be reported
ID kontrolního zvířete – opakování 2
Ammonia levels should be measured immediately prior to renewal
ID exponovaného zvířete – opakování 2
All water quality parameters listed in Table 1of Appendix 1 should be measured immediately prior to renewal
Celkem:
Renewal period should not exceed 72 hours
Tabulka 8
Appropriate feeding schedule (50 % of the daily food ration of commercial tadpole feed)
Zmenšení oblasti folikulárních buněk
Valid experiment in a test determined to be negative for thyroid activity:
Tabulka 9
For any given treatment (including controls), mortality cannot exceed 10 %.
Slovní popis histopatologických nálezů
For any given replicate, mortality cannot exceed three tadpoles, otherwise the replicate is considered compromised
Slovní popis
At least two treatment levels, with all four uncompromised replicates, should be available for analysis
Tabulka 10
At least two treatment levels without overt toxicity should be available for analysis
Šablona souhrnné vykazovací tabulky za den × (7 nebo 21) zkoušky metamorfózy obojživelníků
Valid experiment in a test determined to be positive for thyroid activity:
Kontrola
Mortality of no more than two tadpoles/replicate in the control group can occur
Dávka 1
Decision logic for the conduct of the AMA
Dávka 2
Decision logic was developed for the AMA to provide logical assistance in the conduct and interpretation of the results of the bioassay (see flow chart in Figure 3).
Dávka 3
The decision logic, in essence, weighs the endpoints in that advanced development, asynchronous development and thyroid histopathology are weighed heavily, while delayed development, snout-vent length and wet body weight, parameters that can potentially be affected by general toxicity, are weighed less heavily.
Ukazatel
Advanced development (determined using developmental stage, SVL and HLL)
REP
Advanced development is only known to occur through effects which are thyroid hormone related.
Průměr
These may be peripheral tissue effects such as direct interaction with the thyroid hormone receptor (such as with T4) or effects which alter circulating thyroid hormone levels.
Střed:
In either case, this is considered sufficient evidence to indicate that the chemical has thyroid activity.
Tělesná hmotnost za vlhka
Advanced development is evaluated in one of two ways.
Tabulka 11
First, the general developmental stage can be evaluated using the standardised approach detailed in Nieuwkoop and Faber (8).
Šablona souhrnné vykazovací tabulky údajů o stadiu vývoje za den × (7 nebo 21) zkoušky metamorfózy obojživelníků
Second, specific morphological features may be quantified, such as hind limb length, at both days 7 and 21, which is positively associated with agonistic effects on the thyroid hormone receptor.