natgillin/translations-raw huggingface dataset:
The specific reason for conducting a test will determine the route of exposure (i.e. via water or sediment).
261-287.
Before use of this test method for the testing of a mixture intended for a regulatory purpose, it should be considered whether, and if so why, it may provide adequate results for that purpose.
, Industrial Genotoxicology Group collaborative trial to investigate cell cycle parameters in human lymphocyte cytogenetics studies, Mutagenesis, Vol.12/3, p.163-167.
The test is designed to assess chemical-related effects on the vegetative growth of Myriophyllum plants growing in standardised media (water, sediment and nutrients).
Ames, B.N., J. McCann, E. Yamasaki (1975), Methods for Detecting Carcinogens and Mutagens with the Salmonella/Mammalian Microsome Mutagenicity Test, Mutation Research/Environmental Mutagenesis and Related Subjects, Vol.
For this purpose, shoot apices of healthy, non-flowering plants are potted in standardised, artificial sediment, which is supplemented with additional nutrients to ensure adequate plant growth, and then maintained in Smart and Barko medium (Appendix 1).
31/6, p.
After an establishment period to allow for root formation, plants are exposed to a series of test concentrations added to the water column.
347-363.
Alternatively, exposure via the sediment may be simulated by spiking the artificial sediment with the test chemical and transplanting plants into this spiked sediment.
Ames (1983), Revised Methods for the Salmonella Mutagenicity Test, Mutation Research/Environmental Mutagenesis and Related Subjects, Vol.
In both cases, plants are subsequently maintained under controlled environmental conditions for 14 days.
113/3-4, p.
To quantify chemical-related effects, growth in the test solutions is compared with the growth of the control plants, and the concentration causing a specified x % inhibition of growth is determined and expressed as the ECx; “x” can be any value depending on the regulatory requirements, e.g. EC10 EC20 and EC50.
, Cytogenetic Effects of Mutagens/Carcinogens after Activation in a Microsomal System In Vitro, I. Induction of Chromosomal Aberrations and Sister Chromatid Exchanges by Diethylnitrosamine (DEN) and Dimethylnitrosamine (DMN) in CHO Cells in the Presence of Rat-Liver Microsomes, Mutation Research, Vol.
Both average specific growth rate (estimated from assessments of shoot length, shoot fresh weight and shoot dry weight) and yield (estimated from the increase in shoot length, shoot fresh weight and shoot dry weight) of untreated and treated plants should be determined.
37/1, p. 8390.
If required, the lowest observed effect concentration (LOEC) and the no observed effect concentration (NOEC) may be statistically determined from estimates of average specific growth rates and yield.
Ishidate (1979), Chromosomal Aberration Tests on 29 Chemicals Combined with S9 Mix in vitro, Mutation Research/Genetic Toxicology, Vol.
An analytical method with adequate sensitivity for quantification of the chemicals in the test medium should be available.
66/3, p.
Information on the test chemical which may be useful in establishing the test conditions includes the structural formula, composition in the case of multi-constituent substances, UVCBs, mixtures or formulations, purity, water solubility, stability in water and light, acid dissociation constant (pKa), partition coefficient octanol-water (Kow), if available Kd to sediments, vapour pressure and biodegradability.
277-290.
Water solubility and vapour pressure can be used to calculate Henry's Law constant, which will indicate whether significant losses of the test chemical during the test period are likely.
, Differential effects of cytochrome P450-inducers on promutagen activation capabilities and enzymatic activities of S-9 from rat liver, Journal of Environmental Pathology and Toxicology, Vol.
If losses of the test chemicals are likely, the losses should be quantified and the subsequent steps to control such losses should be documented.
4/1, p.
Where information on the solubility and stability of the test chemical(s) is uncertain, it is recommended that these properties are assessed under the conditions of the test, i.e. growth medium, temperature, lighting regime to be used in the test.
55-65.
Note: when light dependent peroxidising herbicides are tested, the laboratory lighting used should contain the equivalent presence of solar ultraviolet light found in natural sunlight.
Alternatives to Aroclor 1254-induced S9 in in vitro Genotoxicity Assays, Mutagenesis, Vol.
The pH should be measured and adjusted in the test medium as appropriate.
7/3, p.
The pH control of the test medium is particularly important, e.g. when testing metals or chemicals which are hydrolytically unstable.
175-177.
Further guidance for testing chemicals with physical-chemical properties that make them difficult to test is provided in a OECD Guidance Document (11).
Report from Working Group on in vitro Tests for Chromosomal Aberrations, Mutation Research/Environmental Mutagenesis and Related Subjects, Vol.
For the test results to be valid, the mean total shoot length and mean total shoot fresh weight in control plants at least double during the exposure phase of the test.
312/3, p.
In addition, control plants must not show any visual symptoms of chlorosis and should be visibly free from contamination by other organisms such as algae and/or bacterial films on the plants, at the surface of the sediment and in the test medium.
241-261.
The mean coefficient of variation for yield based on measurements of shoot fresh weight (i.e. from test initiation to test termination) in the control cultures does not exceed 35 % between replicates.
Galloway (1996), Human liver S-9 metabolic activation: proficiency in cytogenetic assays and comparison with phenobarbital/beta-naphthoflavone or Aroclor 1254 induced rat S-9, Environmental and Molecular Mutagenesis, Vol.
A reference chemical(s), such as 3,5-dichlorophenol used in the ring test (9), should be periodically tested in order to check the performance of the test procedure over time.
28/1, p.
The ring test data indicate that the mean EC50 values of 3,5-DCP for the different response variables were between 4,7 and 6,1 mg/l (see the ring-test report for details of anticipated confidence interval around these values).
51-59.
It is advisable to test a reference chemical at least twice a year or, where testing is carried out infrequently, in parallel with the definitive toxicity tests.
Christensen (1987), Effects of anticoagulants upon sister-chromatid exchanges, cell-cycle kinetics, and mitotic index in human peripheral lymphocytes, Mutation Research, Vol.
A guide to expected EC50 values for 3,5-DCP is provided in the Statistical Report of the International Ring-test (9).
190/3, p.
Test apparatus
225-8.
The test should be conducted under controlled environmental conditions, i.e. in a growth chamber, growth room or laboratory, with controllable day length, lighting and temperature (see section “Test conditions”, paragraphs 56-58).
, Evaluation of an Exposure System Using Cells Grown on Collagen Gels for Detecting Highly Volatile Mutagens in the CHO/HGPRT Mutation Assay, Environmental and Molecular Mutagenesis, Vol.
Stock cultures should be maintained separately from test vessels.
5/6, p.
The study should be conducted using glass test vessels such as aquaria or beakers; 2-l glass beakers (approximately 24 cm high and 11 cm in diameter) are commonly used.
795-801.
However, other (i.e. larger) vessels may be suitable provided that there is sufficient depth of water to allow unlimited growth and keep the plants submerged throughout the test duration.
, An improved system for exposure of cultured mammalian cells to gaseous compounds in the chromosomal aberration assay, Mutation Research, Vol.
Plastic or glass plant pots (approximately 9 cm diameter and 8 cm high and 500 ml volume) may be used as containers for potting the plants into the sediment.
652/2, p.
Alternatively, glass beakers may be used and are preferred in some cases (e.g. testing hydrophobic chemicals or chemicals with high Kow).
122-130.
The choice of pot/beaker size needs to be considered alongside the choice of test vessels and the preferred test design (see below).
I. Theoretical aspects, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol.
If using Test Design A (one shoot per pot with three pots per vessel) then smaller pots or larger vessels may be needed.
655/1-2, p.
If using Test Design B (three shoots per pot and one pot per vessel) then the stated pot and vessel sizes should be adequate.
1-3.
In all cases, the minimum overlaying water depth should be 12 cm above the top of the sediment and the ratio of sediment surface area/volume to water surface area/volume should be recorded.
, Workshop summary: Top concentration for in vitro mammalian cell genotoxicity assays; and Report from working group on toxicity measures and top concentration for in vitro cytogenetics assays (chromosome aberrations and micronucleus), Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol.
The general approaches described in this test method can be used to test a range of aquatic plant species.
723/2, p.
However, the conditions outlined in this test method have been tailored for testing the water milfoil species, Myriophyllum spicatum.
77-83.
This species belongs to the dicotyledonous family, Haloragaceae.
Honma, M. (2011), Cytotoxicity measurement in in vitro chromosome aberration test and micronucleus test, Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, Vol.
Myriophyllum spicatum (Eurasian water milfoil) is a submerged, rooted species which tolerates a wide range of conditions and is found in both static and flowing water bodies.
724/1-2, p.
M. spicatum is a perennial which dies back to the roots over winter.
86-87.
Plants usually flower and set seed freely although vegetative propagation from axillary buds or stem fragments that detach naturally or after disturbance, is often the primary method of colonisation.
Morita, T., M. Honma, K. Morikawa (2012), Effect of reducing the top concentration used in the in vitro chromosomal aberration test in CHL cells on the evaluation of industrial chemical genotoxicity, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol.
Cultivation of the test organism
741/1-2, p.
Plants may be obtained from natural populations or via aquatic plant suppliers.
32-56.
In both cases, the source of the plants should be documented and species identity should be verified.
Levy (2013), Evaluation of the Highest Concentrations Used in the In Vitro Chromosome Aberrations Assay, Environmental and Molecular Muagenesis, Vol.
If in doubt, use of verified laboratory cultures from known sources is recommended.
54/1, p.
Plants that have been exposed to any chemical contaminants, or collected from sites known to be contaminated, should not be used in this test.
36-43.
In regions where M. spicatum is not readily available during the winter months, long-term maintenance of stock cultures may be necessary under glasshouse or laboratory conditions.
EPA, Office of Chemical Safety and Pollution Prevention (2011), Chemical Substances of Unknown or Variable Composition, Complex Reaction Products and Biological Materials: UVCB Substances,http://www.epa.gov/opptintr/newchems/pubs/uvcb.txt.
Stock cultures should be maintained under conditions similar to the test conditions although irradiance and temperature may be reduced in order to reduce the frequency of culture maintenance (e.g. when Myriophyllum tests are not planned for a period).
, Compilation and use of genetic toxicity historical control Data, Mutation Research, Vol.
Use of larger aquaria and plant pots, than would be used in tests, is recommended in order to allow space for proliferation.
723/2, p.
Sediment and water-media composition should be the same as would be used for tests although alternative methods of sediment fertilisation may be adopted (e.g. use of commercial slow-release fertiliser formulations)
87-90.
Stock plants should be visibly free of contamination with any other organisms, including snails, filamentous algae, fungi and insects, e.g. eggs or larvae of the moth Paraponyx stratiotata and larve or adults of the curculionidae Eubrychius velutus.
Fleiss, J. L., B. Levin, M.C. paik (2003).
Stock cultures should be monitored and transplanted as necessary to avoid development of algal and bacterial contamination.
2946.
Aeration of stock cultures may be beneficial should algal or bacterial contamination become problematic.
Locke-Huhle, C. (1983), Endoreduplication in Chinese hamster cells during alpha-radiation induced G2 arrest, Mutation Research, Vol.
In all cases, plants are cultured/ acclimatised under conditions that are similar, but not necessarily identical, to those used in the test for an adequate period (i.e. > 2 weeks) before their use in a test.
119/3, p.
Flowering stock cultures should not be used in a test as vegetative growth rates generally decline during and after flowering.
403-413.
Sediment
Trosko (1983), Aphidicolin – induced endoreduplication in Chinese hamster cells, Cancer Research, Vol.
The following formulated sediment, based on the artificial sediment used in Chapter C.28 of this Annex (8), is recommended for use in this test.
43/3, p. 1362-1364.
The sediment is prepared as described in TM C.28, except for the addition of nutrients as described below:
Galloway (1994), Cytotoxicity measurement in in vitro chromosome aberration test and micronucleus test, Mutation Research, Vol. 312, p.
4-5 % peat (dry weight, according to 2 ± 0,5 % organic carbon) as close to pH 5,5 to 6,0 as possible; it is important to use peat in powder form, finely ground (preferably particle size < 1 mm) and only air dried.
139-149.
20 % (dry weight) kaolin clay (kaolinite content preferably above 30 %).
Pentru testele prevăzute în această metodă de testare utilizând linii celulare, citotoxicitatea este identificată ca o reducere a dublării relative a populației (RPD) sau creșterea relativă a numărului de celule (RICC) a celulelor tratate în comparație cu martorul negativ (a se vedea punctul 17 și apendicele 2).
75-76 % (dry weight) quartz sand (fine sand should predominate with more than 50 % of the particles between 50 and 200 μm).
pentru studiile pe termen scurt: greutatea corporală a fiecărui animal la începutul și sfârșitul testului; pentru studiile cu durata mai mare de o săptămână: greutatea corporală a fiecărui animal pe durata studiului și consumul de hrană.
An aqueous nutrient medium is added such that the final sediment batch contains 200 mg/Kg dry sediment of both ammonium chloride and sodium phosphate and the moisture content of the final mixture is in a range of 30-50 %.
Se includ intervalul de greutate, valoarea medie și deviația standard pentru fiecare grup.
Calcium carbonate of chemically pure quality (CaCO3) is added to adjust the pH of the final mixture of the sediment to 7,0 ± 0,5.
Analysis of chromosome aberrations in bone marrow cells, Mutation Research, Vol. 189/2, p.
The source of peat, kaolin clay and sand should be known and documented.
157-165.
If the origin is unknown or gives some level of concern, then the respective components should be checked for the absence of chemical contamination (e.g. heavy metals, organochlorine compounds, organophosphorous compounds).
, Report from the working group on the in vivo mammalian bone marrow chromosomal aberration test, Mutation Research, Vol.
The dry constituents of the sediment should be mixed homogenously prior to mixing the aqueous nutrient solution thoroughly into the sediment.
312/3, p.
The moist sediment should be prepared at least two days before use to allow thorough soaking of the peat and to prevent hydrophobic peat particles floating to the surface when the sediment is overlaid with media; before use, the moist sediment may be stored in the dark.
305-312.
For the test, the sediment is transferred into a suitable size containers, such as plant pots of a diameter which fit into the glass vessels (the sediment surface area should cover approximately 70 % or more of the vessel surface area).
, Recommendations for statistical designs of in vivo mutagenicity tests with regard to subsequent statistical analysis, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol.
In cases where the container has holes at the bottom, a piece of filter paper in the bottom of the container will help to keep the sediment within the container.
417/1, p.
The pots are filled with the sediment such that the sediment surface is level, prior to covering with a thin layer (~ 2 to 3 mm) of an inert material such as sand, fine horticultural grit (or crushed coral) to keep the sediment in place.
19-30.
Smart and Barko medium (12) is recommended for culturing and testing Myriophyllum spicatum.
, Compilation and use of genetic toxicity historical control data, Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, Vol.
Preparation of this media is described in the Appendix 1.
723/2, p.
The pH of the media (water phase) at test initiation should be between 7,5 and 8,0 for optimum plant growth.
87-90.
Experimental design
, in vivo rodent erythrocyte micronucleus assay, Mutation Research/Environmental Mutagenesis and Related Subjects, Vol. 312/3, p.
The test should incorporate a minimum of six replicate test vessels for the untreated control and a minimum of four replicate test vessels for each of a minimum of five concentration levels.
293-304.
Each test vessel represents a replicate containing three shoots.
Dose setting in in vivo mutagenicity assays, Mutagenesis, Vol.
There are two options for growing three shoots in each test vessel:
7/5, p.
Test Design A: one shoot per pot and three pots per vessel.
313-319.
Test Design B: three shoots per pot and one pot per vessel.
Pacchierotti, F., V. Stocchi (2013), Analysis of chromosome aberrations in somatic and germ cells of the mouse, Methods in Molecular Biology, Vol. 1044, p.
Alternative test designs of one shoot per pot per test vessel are acceptable provided that replication is adjusted as required to achieve the required validity criteria.
147-163.
The individual test vessels should be randomly assigned to the treatment groups.
pentru studiile pe termen scurt: greutatea corporală a fiecărui animal la începutul și la sfârșitul testului; pentru studiile mai lungi de o săptămână: greutatea corporală a fiecărui animal pe durata studiului și consumul de hrană.
A randomised design for the location of the test vessels in the test area is required to minimise the influence of spatial differences in light intensity or temperature.
Validation and regulatory acceptance of automated scoring and the use of rat peripheral blood reticulocytes, with discussion of non-hematopoietic target cells and a single dose-level limit test, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol.
Test chemical concentrations and control groups
627/1, p.
To determine an ECx, test concentrations should bracket the ECx to ensure an appropriate level of confidence.
94/1, p.
The use of more test concentrations will improve the confidence interval around the resulting ECx value.
92-107.
To determine the LOEC/NOEC (optional endpoint), the lowest test concentration should be sufficiently low such that growth is not significantly different from growth in control plants.
, Flow cytometric analysis of micronuclei in peripheral blood reticulocytes: I. Intra- and interlaboratory comparison with microscopic scoring, Toxicological Sciences, Vol.
In addition, the highest test concentration should be sufficiently high such that growth is significantly lower than that in the control.
94/1, p.
The use of more replicates will enhance the statistical power of the no effect-concentration/ ANOVA design.
83-91.
In cases where a range-finding test indicates that the test chemical does not have an adverse effect at concentrations up to 100 mg/l or up to its limit of solubility in the test medium, or in the case of a formulation up to the limit of dispersibility, a limit test may be undertaken to facilitate comparison of responses in a control group and one treatment group — 100 mg/l or a concentration equal to the limit of solubility, or 1000 mg/kg dry sediment.
, Flow cytometric scoring of micronucleated erythrocytes: an efficient platform for assessing in vivo cytogenetic damage, Mutagenesis, Vol.
This test should follow the general principles of a standard dose-response test, with the exception that an increase in the minimum number of replicates to six test vessels per control and concentration is advised.
26/1, p.
Test solutions are usually prepared by dilution of a stock solution, prepared by dissolving or dispersing the test chemical in Smart and Barko media, using demineralised (i.e. distilled or deionised) water (see Appendix 1).
139-145.
The highest test concentration should normally not exceed the water solubility of the test chemical or, in the case of formulations, the dispersibility under the test conditions.
Garriott (1996), Validation of an automated image analysis micronucleus scoring system, Mutation Research, Vol.
For test chemicals of low water solubility, it may be necessary to prepare a concentrated stock solution or dispersion of the chemical using an organic solvent or dispersant in order to facilitate the addition of accurate quantities of the test chemical to the test medium and aid in its dispersion and dissolution.
370/1, p.
Every effort should be made to avoid the use of such solvents or dispersants.
65-73.
For example, commonly used solvents, which do not cause phytotoxicity at concentrations up to 100 μl/l, include acetone and dimethylformamide.
, An automated new technique for scoring the rodent micronucleus assay: computerized image analysis of acridine orange supravitally stained peripheral blood cells, Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, Vol.
If a solvent or dispersant is used, its final concentration should be reported and kept to a minimum (≤ 100 μl/l).
404/1-2, p.
Under these circumstances all treatments and (solvent) controls should contain the same concentration of solvent or dispersant.
149-154.
Untreated control replicates that do not contain a solvent or dispersant are also incorporated into the test design.
, Automation of mouse micronucleus genotoxicity assay by laser scanning cytometry, Cytometry, Vol.
Further guidance on the use of dispersants is given in an OECD Guidance Document (11).
44/2, p.
The test procedure varies according to the application route of the test chemical (i.e. via the water or sediment phase).
153-155.
The likely behaviour of the test chemical in a water-sediment system should be considered to inform the choice of exposure regime used in the test (i.e. static or static renewal, spiked water or spiked sediment).
, A rapid in vivo test for chromosomal damage, Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, Vol. 18/2, p.
Spiked sediment tests may be preferred in some cases for chemicals that are predicted to significantly partition to sediment.
187-190.
Establishment phase
, The micronucleus test, Mutation Research, Vol.
Healthy shoot apices/tips, i.e. without side shoots, are cut from the culture plants to give a shoot length of 6 cm (± 1 cm).
31/1, p. 9-15.
For Test Design A (one shoot per pot and three pots per vessel) single shoot tips are planted into each pot.
Environmental Protection Agency Gene-Tox Program, Mutation Research/Reviews in Genetic Toxicology, Vol. 123/1, p.
For Test Design B (three shoots per pot and one pot per vessel) four to five shoot apices are planted into each pot containing the sediment.
61-118.
In both cases surplus pots should be planted to allow for selection of uniform plants at test initiation, as well as to provide spare plants to be used for inspection of root growth immediately prior to treatment and spare plants to be harvested for shoot biomass and length measurements on Day 0.
Environmental Protection Agency Gene-Tox Program, Mutation Research/Reviews in Genetic Toxicology, Vol.
Shoots are inserted such that approximately three cm, covering at least two nodes, are beneath the sediment surface.
239/1, p.