natgillin/translations-raw huggingface dataset:
Following a 48-hour settling-in period, mortalities are recorded and the following criteria applied:
Količina preskusne raztopine
mortalities of greater than 10 % of population in seven days:
8 l (minimum)
reject the entire batch;
1,5 l (minimum)
mortalities of between 5 % and 10 % of population:
4 l (minimum)
acclimation for seven additional days;
Zamenjave prostornin preskusnih raztopin
if more than 5 % mortality during second seven days, reject the entire batch;
vsaj 6 na dan
mortalities of less than 5 % of population in seven days:
vsaj 5 na dan
accept the batch
glej odstavek 20
Fish should not receive treatment for disease during the acclimation period, in the pre-exposure period, or during the exposure period.
1,5 ± 20 %
Pre-exposure and selection of fish
samci:
A one-week pre-exposure period is recommended, with animals placed in vessels similar to the actual test.
2,5 ± 20 %
Fish should be fed ad libitum throughout the holding period and during the exposure phase.
samice:
The exposure phase is started with sexually dimorphic adult fish from a laboratory supply of reproductively mature animals (e.g. with clear secondary sexual characteristics visible as far as fathead minnow and medaka are concerned), and actively spawning.
0,35 ± 20 %
For general guidance only (and not to be considered in isolation from observing the actual reproductive status of a given batch of fish), fathead minnows should be approximately 20 (± 2) weeks of age, assuming they have been cultured at 25 ± 2 °C throughout their lifespan.
0,65 ± 20 %
Japanese medaka should be approximately 16 (± 2) weeks of age, assuming they have been cultured at 25 ± 2 °C throughout their lifespan.
0,4 ± 20 %
Zebrafish should be approximately 16 (± 2) weeks of age, assuming they have been cultured at 26 ± 2 °C throughout their lifespan.
Št. rib na preskusno posodo
TEST DESIGN
6 (2 samca in 4 samice)
Three concentrations of the test chemical, one control (water) and, if needed, one solvent control are used.
10 (5 samcev in 5 samic)
The data may be analysed in order to determine statistically significant differences between treatment and control responses.
Št. tretiranj
These analyses will inform whether further longer term testing for adverse effects (namely, survival, development, growth and reproduction) is required for the chemical, rather than for use in risk assessment (24).
= 3 (plus ustrezno št. kontrol)
For zebrafish and medaka, on day 21 of the experiment, males and females from each treatment level (5 males and 5 females in each of the two replicates) and from the control(s) are sampled for the measurement of vitellogenin and secondary sex characteristics, where applicable.
Št. posod na tretiranje
For fathead minnow, on day 21 of exposure, males and females (2 males and 4 females in each of the four replicates) and from the control(s) are sampled for the measurement of vitellogenin and secondary sex characteristics.
najmanj 4
For the purposes of this test, the highest test concentration should be set by the maximum tolerated concentration (MTC) determined from a range finder or from other toxicity data, or 10 mg/l, or the maximum solubility in water, whichever is lowest.
najmanj 2
The MTC is defined as the highest test concentration of the chemical which results in less than 10 % mortality.
Št. rib na preskusno koncentracijo
Using this approach assumes that there are existing empirical acute toxicity data or other toxicity data from which the MTC can be estimated.
16 odraslih samic in 8 samcev (4 samice in 2 samca v vsaki ponovitveni posodi)
Estimating the MTC can be inexact and typically requires some professional judgment.
10 odraslih samic in 10 samcev (5 samic in 5 samcev v vsaki ponovitveni posodi)
Three test concentrations, spaced by a constant factor not exceeding 10, and a dilution-water control (and solvent control if necessary) are required.
Način hranjenja
A range of spacing factors between 3,2 and 10 is recommended.
živi ali zamrznjeni odrasli solinski rakci ali njihovi navpliji dvakrat ali trikrat na dan (ad libitum), komercialno dostopna hrana ali kombinacija navedenega
Selection and weighing of test fish
navpliji solinskih rakcev dvakrat ali trikrat na dan (ad libitum), komercialno dostopna hrana ali kombinacija navedenega
It is important to minimise variation in weight of the fish at the beginning of the assay.
brez, razen če koncentracija raztopljenega kisika pade pod 60 % nasičenosti
Suitable size ranges for the different species recommended for use in this test are given in Appendix 2.
čista površinska voda, voda iz vodnjaka ali modelna razredčevalna voda ali deklorirana vodovodna voda
For the whole batch of fish used in the test, the range in individual weights for male and female fish at the start of the test should be kept, if possible, within ± 20 % of the arithmetic mean weight of the same sex.
Obdobje predhodne izpostavljenosti
It is recommended to weigh a subsample of the fish stock before the test in order to estimate the mean weight.
priporočeno 7 dni
Conditions of exposure
Trajanje izpostavljenosti kemikaliji
The test duration is 21 days, following a pre-exposure period.
Biološke končne točke
The recommended pre-exposure period is one week.
preživetje
Fish should be fed ad libitum with an appropriate food (Appendix 2) at a sufficient rate to maintain body condition.
obnašanje
Care should be taken to avoid microbial growth and water turbidity.
spolne značilnosti
As a general guidance, the daily ration may be divided into two or three equal portions for multiple feeds per day, separated by at least three hours between each feed.
Sprejemljivost preskusa
A single larger ration is acceptable particularly for weekends.
raztopljeni kisik > 60 % nasičenosti;
Food should be withheld from the fish for 12 hours prior to sampling/necropsy.
srednja temperatura 25 ± 2 °C;
Fish food should be evaluated for the presence of contaminants such as organochlorine pesticides, polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs).
dstotno preživetje rib v kontrolah;
Food with an elevated level of phytoestrogens that would compromise the response of the assay to known oestrogen agonist (e.g. 17-beta estradiol) should be avoided.
izmerjene preskusne koncentracije v razponu 20 % srednjih izmerjenih vrednosti na stopnjo tretiranja
Uneaten food and faecal material should be removed from the test vessels at least twice weekly, e.g. by carefully cleaning the bottom of each tank using a siphon.
srednja temperatura 24 ± 2 °C;
The photoperiod and water temperature should be appropriate for the test species (see Appendix 2).
srednja temperatura 26 ± 2 °C;
Frequency of analytical determinations and measurements
Nekatere kemične značilnosti sprejemljive vode za redčenje
Prior to initiation of the exposure period, proper function of the chemical delivery system should be ensured.
< 20 mg/l
All analytical methods needed should be established, including sufficient knowledge on the chemical stability in the test system.
< 2 mg/l
During the test, the concentrations of the test chemical are determined at regular intervals, as follows:
skupni organofosforni pesticidi
the flow rates of diluent and toxicant stock solution should be checked preferably daily but as a minimum twice per week, and should not vary by more than 10 % throughout the test.
Dodatek 4A
It is recommended that the actual test chemical concentrations be measured in all vessels at the start of the test and at weekly intervals thereafter.
Substrat za drstenje cebrice
It is recommended that results be based on measured concentrations.
Drstilni pladenj
However, if concentration of the test chemical in solution has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test, then the results can either be based on nominal or measured values.
popolnoma steklena laboratorijska posoda, na primer 22 × 15 × 5,5 cm (d × š × g), pokrita z odstranljivo rešetko iz nerjavnega jekla (širina mreže 2 mm).
Samples may need to be filtered (e.g., using a 0,45 μm pore size) or centrifuged.
Rešetka mora prekriti odprtino laboratorijske posode pod robom.
If needed, then centrifugation is the recommended procedure.
Na rešetko je treba pritrditi substrat za drstenje.
However, if the test material does not adsorb to filters, filtration may also be acceptable.
Zagotoviti mora strukturo, v katero se premaknejo ribe.
During the test, dissolved oxygen, temperature, and pH should be measured in all test vessels at least once per week.
Primerne so na primer umetne akvarijske rastline iz zelene plastike (opomba: proučiti je treba morebitno adsorpcijo preskusne kemikalije na plastični material).
Total hardness and alkalinity should be measured in the controls and one vessel at the highest concentration at least once per week.
Plastični material je treba dovolj dolgo izpirati v zadostni količini tople vode za zagotovitev, da v preskusno vodo ne bo prešla nobena kemikalija.
Temperature should preferably be monitored continuously in at least one test vessel.
Pri uporabi steklenih materialov je treba zagotoviti, da se ribe med intenzivnim premikanjem ne poškodujejo niti niso utesnjene.
A number of general (e.g. survival) and core biological responses (e.g. vitellogenin levels) are assessed over the course of the assay or at termination of the assay.
Razdalja med pladnjem in steklenimi ploščami mora biti vsaj 3 cm, da bi bilo zagotovljeno, da se ribe ne drstijo zunaj pladnja.
Measurement and evaluation of these endpoints and their utility are described below.
Ikre, odložene na pladenj, padejo skozi rešetko in se lahko vzorčijo 45–60 minut po začetku osvetljenosti.
Survival
Prozorne ikre niso sprijete in se lahko enostavno štejejo z uporabo prečne svetlobe.
Fish should be examined daily during the test period and any mortality should be recorded and the dead fish removed as soon as possible.
Če je v posodi pet samic, se do 20 iker na dan lahko šteje za nizko število, do 100 iker kot srednje in več kot 100 iker kot visoko število.
Dead fish should not be replaced in either the control or treatment vessels.
Drstilni pladenj je treba odstraniti, zbrati ikre in drstilni pladenj ponovno postaviti v preskusno posodo čim pozneje zvečer ali zelo zgodaj zjutraj.
Sex of fish that die during the test should be determined by macroscopic evaluation of the gonads.
Čas do ponovne postavitve rešetke ne sme presegati eno uro, saj bi sicer lahko substrat za drstenje vodil do posameznih parjenj in drstenj ob neobičajnem času.
Behaviour and appearance
Če je treba zaradi razmer drstilni pladenj vstaviti pozneje, je treba to storiti vsaj 9 ur po začetku osvetljenosti.
Any abnormal behaviour (relative to controls) should be noted;
Drstenje se tako pozno ne začne več.
this might include signs of general toxicity including hyperventilation, uncoordinated swimming, loss of equilibrium, and atypical quiescence or feeding.
Dodatek 4B
Additionally external abnormalities (such as haemorrhage, discoloration) should be noted.
Substrat za drstenje črnoglavega pisanca
Such signs of toxicity should be considered carefully during data interpretation since they may indicate concentrations at which biomarkers of endocrine activity are not reliable.
V vsako preskusno komoro se postavijo dve ali tri kombinirane plastične/keramične/steklene plošče ali plošče iz nerjavnega jekla in drstilni pladnji (npr. 80 mm dolg siv polkrožen žleb na pladenj z robovi, dolg 130 mm) (glej sliko).
Such behavioural observations may also provide useful qualitative information to inform potential future fish testing requirements.
Ustrezno pripravljene plošče iz PVC ali keramike so dokazano primerne za substrat za drstenje (Thorpe idr., 2007).
For example, territorial aggressiveness in normal males or masculinised females has been observed in fathead minnows under androgenic exposure;
Priporočeno je, da so plošče pobrušene, da se izboljša oprijem.
in zebrafish, the characteristic mating and spawning behaviour after the dawn onset of light is reduced or hindered by oestrogenic or anti-androgenic exposure.
Pladenj mora biti tudi zaščiten, da se ribam prepreči dostop do iker, ki padejo skozi, razen če je bil za uporabljen substrat za drstenje dokazan učinkovit oprijem.
Because some aspects of appearance (primarily colour) can change quickly with handling, it is important that qualitative observations be made prior to removal of animals from the test system.
Podlaga je zasnovana tako, da zadrži vse ikre, ki se ne primejo površine plošče in bi zato padle na dno posode (ali tiste ikre, ki so odložene neposredno na ravno plastično podlago).
Experience to date with fathead minnows suggests that some endocrine active chemicals may initially induce changes in the following external characteristics:
Vse substrate za drstenje je treba pred uporabo izpirati v vodi za redčenje vsaj 12 ur.
body colour (light or dark), coloration patterns (presence of vertical bands), and body shape (head and pectoral region).
Dodatek 5A
Therefore observations of physical appearance of the fish should be made over the course of the test, and at conclusion of the study
Ocena sekundarnih spolnih značilnosti pri črnoglavem pisancu za zaznavanje nekaterih kemikalij z endokrinim delovanjem
Humane killing of fish
Pregled
At day 21, i.e. at termination of the exposure, the fish should be euthanized with appropriate amounts of Tricaine (Tricaine methane sulfonate, Metacain, MS-222 (CAS 886-86-2), 100-500 mg/l buffered with 300 mg/l NaHCO3 (sodium bicarbonate, CAS 144-55-8) to reduce mucous membrane irritation; blood or tissue is then sampled for vitellogenin determination, as explained in the Vitellogenin section.
Potencialno pomembne značilnosti fizičnega videza odraslih črnoglavih pisancev pri preskušanju povzročiteljev endokrinih motenj vključujejo barvo telesa (tj. svetlo/temno), vzorce obarvanja (tj. prisotnost navpičnih prog), obliko telesa (tj. oblika glave in območja prsi, napetost trebuha) in posebne sekundarne spolne značilnosti (tj. število in velikost drstnih izpuščajev, velikost dorzalne strani trupa pred hrbtno plavutjo in leglica).
Observation of secondary sex characteristics
Drstni izpuščaji so na glavi (dorzalna stran trupa pred hrbtno plavutjo) razmnoževalno aktivnih samcev črnoglavega pisanca in so običajno razporejeni v dvostransko simetrični vzorec (Jensen idr., 2001).
Some endocrine active chemicals may induce changes in specialised secondary sex characteristics (number of nuptial tubercles in male fathead minnow, papillary processes in male medaka).
Pri kontrolnih samicah ter mladih samcih in samicah razvoj izpuščajev ni viden (Jensen idr., 2001).
Notably, chemicals with certain modes of action may cause abnormal occurrence of secondary sex characteristic in animals of the opposite sex;
Okoli oči in med nosnicama samcev je lahko do osem posameznih izpuščajev.
for example, androgen receptor agonists, such as trenbolone, methyltestosterone and dihydrotestosterone, can cause female fathead minnows to develop pronounced nuptial tubercles or female medaka to develop papillary processes (11, 20, 21).
Največ izpuščajev in največji izpuščaji se nahajajo v dveh vzporednih linijah neposredno pod nosnicama in nad usti.
It also has been reported that oestrogen receptor agonists can decrease nuptial tubercle numbers and size of the dorsal nape pad in adult males (25, 26).
Pri veliko ribah so skupine izpuščajev pod spodnjo čeljustjo;
Such gross morphological observations may provide useful qualitative and quantitative information to inform potential future fish testing requirements.
izpuščaji, ki so najbliže ustom, se običajno pojavljajo kot par, medtem ko lahko bolj ventralen sklop sestavljajo največ štirje izpuščaji.
The number and size of nuptial tubercles in fathead minnow and papillary processes in medaka can be quantified directly or more practically in preserved specimens.
Dejansko število izpuščajev je redko večje od 30 (razpon 18–28; Jensen idr., 2001).
Recommended procedures for the evaluation of secondary sex characteristics in fathead minnow and medaka are available from Appendix 5A and Appendix 5B, respectively.
Prevladujoči izpuščaji (v smislu števila) so prisotni kot posamezne, razmeroma okrogle strukture, katerih višina je približno enaka polmeru.
Vitellogenin (VTG)
Večina razmnoževalno aktivnih samcev ima tudi vsaj nekaj izpuščajev, ki so večji in izraziti, da jih ni mogoče razlikovati kot posamezne strukture.
Blood is collected from the caudal artery/vein with a heparinised microhematocrit capillary tubule, or alternatively by cardiac puncture with a syringe.
Nekatere vrste kemikalij, ki povzročajo endokrine motnje, lahko povzročijo nenormalen pojav nekaterih sekundarnih spolnih značilnosti pri nasprotnem spolu;
Depending upon the size of the fish, collectable blood volumes generally range from 5 to 60 μl per individual for fathead minnows and 5-15 μl per individual for zebrafish.
agonisti androgenih receptorjev, kot je 17β-metiltestosteron ali 17β-trenbolon, lahko pri samicah črnoglavega pisanca povzročijo razvoj drstnih izpuščajev (Smith 1974; Ankley idr., 2001; 2003), medtem ko lahko agonisti estrogenih receptorjev zmanjšajo število ali velikost drstnih izpuščajev pri samcih (Miles-Richardson idr.
Plasma is separated from the blood via centrifugation, and stored with protease inhibitors at – 80 °C, until analysed for vitellogenin.
1999; Harries idr.
Alternatively, in medaka the liver will be used, and in zebrafish the head/tail homogenate can be used as tissue-source for vitellogenin determination (Appendix 6).
2000).
The measurement of VTG should be based upon a validated homologous ELISA method, using homologous VTG standard and homologous antibodies.
V nadaljevanju je naveden opis opredelitve drstnih izpuščajev pri črnoglavem pisancu na podlagi postopkov, uporabljenih v laboratoriju Agencije za varstvo okolja ZDA (U. S. Environmental Protection Agency) v Duluthu, Minnesota.
It is recommended to use a method capable to detect VTG levels as low as few ng/ml plasma (or ng/mg tissue), which is the background level in unexposed male fish.
Specifični izdelki in/ali oprema se lahko nadomestijo s primerljivimi materiali, ki so na voljo.
Quality control of vitellogenin analysis will be accomplished through the use of standards, blanks and at least duplicate analyses.
Opazovanje je najboljše z uporabo osvetljenega povečevalnega stekla ali trikratno osvetljenega stereomikroskopa.
For each ELISA method, a test for matrix effect (effect of sample dilution) should be run to determine the minimum sample dilution factor.
Ribe se opazujejo dorzalno in anteriorno naprej (glava proti opazovalcu).
Each ELISA plate used for VTG assays should include the following quality control samples:
Ribe se postavijo v majhno petrijevko (npr. s premerom 100 mm) anteriorno naprej in ventralno navzdol.
at least 6 calibration standards covering the range of expected vitellogenin concentrations, and at least one non-specific binding assay blank (analysed in duplicate).
Iskalo se usmeri tako, da se omogoči identifikacija izpuščajev.
Absorbance of these blanks should be less than 5 % of the maximum calibration standard absorbance.
Za identifikacijo območij z izpuščaji se riba nežno in počasi obrne z ene strani na drugo.
At least two aliquots (well-duplicates) of each sample dilution will be analysed.
Izpuščaji se preštejejo in ocenijo.
Well-duplicates that differ by more than 20 % should be re-analysed.
Opazovanje se ponovi na ventralni površini glave, tako da se riba v petrijevko postavi dorzalno anteriorno naprej.
If the nominal concentrations trend away from the calibration regression line (e.g. at lower concentrations), it may be necessary to split the calibration curve into low and high ranges or to use a nonlinear model to adequately fit the absorbance data.
Za določitev lokacije in količine prisotnih izpuščajev je bila pripravljena predloga (glej konec tega Dodatka).
The ratio of the expected concentration to the measured concentration will be reported along with the results from each set of assays performed on that day.
Ocena 1 – prisoten, ta ocena se opredeli kot vsak izpuščaj z eno točko, v kateri je višina skoraj enaka polmeru (premeru).
Evaluation of Biomarker Responses by Analysis of Variance (ANOVA)
Ocena 2 – povečan, ta ocena se opredeli s tkivom, ki je po videzu podobno zvezdici in ima običajno veliko zvezdasto osnovo z brazdami ali gubami, ki izhajajo in središča.
To identify potential endocrine activity of a chemical, responses are compared between treatments and control groups using analysis of variance (ANOVA).
Izpuščaj je v višino pogosto bolj žagast, vendar je lahko občasno nekoliko zaokrožen.
Where a solvent control is used, an appropriate statistical test should be performed between the dilution water and solvent controls for each endpoint.
Ocena 3 – izrazit, ta ocena običajno pomeni razmeroma velik in zaokrožen izpuščaj z manj definirano strukturo.
Guidance on how to handle dilution water and solvent control data in the subsequent statistical analysis can be found in OECD, 2006c (27).
Ti izpuščaji se občasno nahajajo skupaj in tvorijo eno maso skupaj s posameznimi območji (B, C in D, opisana v nadaljevanju) ali njihovo kombinacijo.
All biological response data should be analysed and reported separately by sex.
Obarvanost in oblika sta podobna oceni 2, vendar sta občasno razmeroma neselektivna.
If the required assumptions for parametric methods are not met — non-normal distribution (e.g. Shapiro-Wilk's test) or heterogeneous variance (Bartlett's test or Levene's test), consideration should be given to transforming the data to homogenise variances prior to performing the ANOVA, or to carrying out a weighted ANOVA.
S tem ocenjevalnim sistemom je skupna ocena izpuščajev običajno < 50 pri normalnih kontrolnih samcih s številom izpuščajev od 18 do 20 (Jensen idr., 2001).
Dunnett's test (parametric) on multiple pair-wise comparisons or a Mann-Whitney with Bonferroni adjustment (non-parametric) may be used for non-monotonous dose-response.
Dejansko število izpuščajev pri nekaterih ribah je lahko večje od polj v predlogi (Dodatek A) za določeno območje ocenjevanja.
Other statistical tests may be used (e.g. Jonckheere-Terpstra test or Williams test) if the dose-response is approximately monotone.
V tem primeru se lahko označijo dodatne ocene, in sicer desno ali levo od polja.
A statistical flowchart is provided in Appendix 8 to help in the decision on the most appropriate statistical test to be used.
Zato ni treba, da je predloga simetrična.
Additional information can also be obtained from the OECD Document on Current Approaches to Statistical Analysis of Ecotoxicity Data (27).
Dodatna tehnika označevanja izpuščajev, ki so v parih ali združeni navpično ob vodoravni ravnini ust, se lahko izvede z označevanjem dveh ocen izpuščajev v enem polju.
Reporting of test results
Območja označevanja:
Study data should include:
A izpuščaji okoli oči.
Responsible personnel and their study responsibilities
Pri odraslih kontrolnih samcih jih je pogosto več, pri kontrolnih samicah jih ni, na splošno so v parih (en blizu vsakega očesa) ali sami pri samicah, izpostavljenih androgenom.
Each laboratory should have demonstrated proficiency using a range of representative chemicals
B izpuščaji med nosnicama (pore senzornega kanala).
Characterisation of test chemical
Pri kontrolnih samcih so običajno v parih in v razvitejših fazah (2 – povečani ali 3 – izraziti).
Physical nature and relevant physicochemical properties
Niso prisotni pri kontrolnih samicah, pri čemer se pojavljajo in razvijajo pri samicah, izpostavljenih androgenom.
Solvent:
Pri manj razvitih samcih ali samicah, tretiranih z androgenom, so prisotni ali povečani.
Characterization of solvent (nature, concentration used)
D izpuščaji vzporedno z ustno linijo.
Justification of choice of solvent (if other than water)
Pri kontrolnih samcih so običajno ocenjeni kot razviti.
Test animals:
Pri kontrolnih samicah niso prisotni, vendar so prisotni pri samicah, izpostavljenih androgenom.
Species and strain
E izpuščaji na spodnji čeljusti, blizu ust, so običajno majhni in pogosto v parih.
Supplier and specific supplier facility
Pri kontrolnih ali tretiranih samcih in tretiranih samicah se razlikujejo.
Details of animal acclimation procedure
Prisotni so pri kontrolnih samcih in samicah, izpostavljenih androgenom.