Chemicals—Test method of sex-linked recessive lethal in Drosophila Melanogaster

GB/T 27822-2011 Test Method for Sex-Linked Recessive Lethality of Drosophila melanogaster in Chemicals GB/T27822-2011 Standard compression package decompression password: www.bzxz.net
This standard specifies the terms and definitions, reference materials, basic test principles, test methods, test data and reports for the sex-linked recessive lethality test of Drosophila melanogaster. This standard is applicable to the sex-linked recessive lethality test of Drosophila melanogaster for the detection of chemicals. This
standard was drafted in accordance with the plan given in GB/T1.1-2009. This
standard is consistent with the technical content of the Organization for Economic Cooperation and Development (OECD) Chemical Test Method No. 477 (1984) "Sex-Linked Recessive Lethality Test for Drosophila melanogaster" (English version).
This standard has been modified in the following structural and editorial aspects:
———A chapter on scope has been added;
———The “reference materials” section in the original text of OECD477 is adopted as Chapter 3 of this standard;
———The “overview of test operation” section in the original text of OECD477 is adopted as Section 5.1 of this standard;
———The “essential information” section in the original text of OECD477 is adopted as Section 5.2 of this standard;
———The measurement units are changed to the legal measurement units of China.
This standard is proposed and managed by the National Technical Committee for Standardization of Dangerous Chemicals Management (SAC/TC251).
The drafting units of this standard are: Institute of Occupational Health and Poisoning Control, Chinese Center for Disease Control and Prevention, Tianjin Center for Disease Control and Prevention, and China Chemical Economic and Technological Development Center.
The main drafters of this standard.

1CS 13. 300,11. 100
National Standard of the People's Republic of China
GB/T27822—2011
Chemicals
Test method of sex-linked recessive lethal in Drosophila Melanogaster2011-12-30Issued
General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of ChinaStandardization Administration of the People's Republic of China
2012-08-01Implementation
This standard was drafted in accordance with the plan given in GB/T 1.1-2009. 27822---2011
This standard is consistent with the technical content of the Organization for Economic Cooperation and Development (OECD) Chemical Test Method No. 477 (1981) Drosophila melanogaster Sex-Linked Recessive Lethal Test. This standard has made the following structural and editorial changes: - Added a chapter on scope; - The "reference materials" part of the original OECD47? is used as Chapter 3 of this standard; - The "overview of test operation" part of the original OECD477 is used as Section 5.1 of this standard; - The "essential information" part of the original OECD477 is used as Section 5.2 of this standard; - The measurement unit is changed to the legal measurement unit of my country. This standard is proposed and managed by the National Technical Committee for Hazardous Chemicals Management Standardization (SAC/TC25I). Drafting of this standard Units: Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention, Tianjin Center for Disease Control and Prevention, China Chemical Economic and Technological Development Center.
The main drafters of this standard: Wu Weigui, Li Chaolin, Wang Yanrang, Wang Xiaobing, Yang De, Liu Jing, Liu Baofeng, Zhang Ming. I
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1 Scope
Test method for sex-linked recessive lethality of Drosophila melanogaster in chemicals GB/T27B22—2011
This standard specifies the terms and definitions, reference materials, basic principles of the test, test methods, test data and reports for the sex-linked recessive lethality test of Drosophila melanogaster.
This standard is applicable to the sex-linked recessive lethality test of Drosophila aeruginosa for the detection of chemicals. 2 Terms and definitions
The following terms and definitions apply to this document. 2. 1
lethal mutation
a change in the genome that causes death in the carrier when expressed. 2.2
recessive mutation
a change in the genome that is expressed under homozygous or hemizygous conditions. 2.3Www.bzxZ.net
sex-linked genes
genes present on sex chromosomes (X or Y), in this method only refers to genes located on the X chromosome. 3 Reference substances
Substances that can be used as positive controls include: ethyl methanesulphonate, N-nitrosodimethylamine.
4 Basic principles of the test
The sex-linked recessive letha (SLRL) test uses Drosophila Melanogaster to detect the incidence of mutations in the insect germ cell line, including point mutations and small deletions. This method is a forward mutation analysis method that can screen mutations in 800 gene loci on the X chromosome, that is, 80% of all X chromosome gene loci. The X chromosome accounts for about 1/51 of the entire genome
Mutations in the X chromosome of Drosophila melanogaster have typical phenotypic manifestations in female individuals carrying the mutant gene. When a hemizygous lethal mutation occurs, it can be inferred by the deletion of one of the two types of female offspring of normal reproduction of heterozygous female individuals. The SLRL test utilizes the characteristics of chromosome-specific markers and arrangements. 5 Test methods
5.1 Overview of test operations
Wild-type male Drosophila melanogaster were exposed to poison and mated with suitable female Drosophila melanogaster. The mated female flies are then mated individually with male TTTKAONTKACA
GB/T27822—2011
Drosophila melanogaster of the same generation, and the phenotype of wild-type eagle-shaped melanogaster fruit in each individual mating offspring is recorded in their next generation. The absence of the wild-type male melanogaster phenotype indicates that the reproductive cells of P1 male agastric fruit flies have undergone sex-linked recessive lethal mutations.
5. 2 Necessary conditions
The necessary conditions for this test are as follows:
Solid, liquid, vapor or gaseous test sample: Chemical identification number of the test sample
-Purity of the chemical (impurities):
-Solubility;
Melting point/boiling point:
-Acidity (measurable)
··Vapor pressure (if known).
5.3 Experimental Animals
5.3.1 Animal Species
Wild-type female and Muller-5-sex Drosophila melanogaster were used. Other female strains with multiple inverted X chromosomes that are appropriately labeled may also be used.
5.3.2 Test Substances
The test substance should be dissolved in water. Substances that are insoluble in water may be dissolved or suspended in a suitable excipient (e.g., a mixture of ethanol and Tween 60 or 80) and then diluted with water or ice before sampling. Avoid using dimethylsulfoxide (DMSO) as an excipient.
5.4 Experimental Conditions
5.4.1, Administration
Administration may be by oral administration, injection or exposure to gas or vapor. Test substances administered orally should be dissolved in sucrose water. If necessary, the test substance can be dissolved in 0.7% Na solution and injected into the thoracic or abdominal cavity. 5.4.2 Dose level
When conducting a preliminary evaluation of the test substance mutation, test a maximum tolerated dose or the highest dose that can be achieved. If the purpose is to determine the dose-response relationship, at least three dose groups are required. 5.4.3 Control
The test should have a negative control group (excipient) and a positive control group. Substances that can be used as positive controls include: ethyl methanesulfonate: N-nitrosodimethylamine. If there are appropriate laboratory control data, no control group is required. 5.5 Test Procedures
To detect the sensitivity of the test substance to different stages of reproductive cells, male flies (3-5 days old) were mated with virgin flies of Muller-5 strain or other strains with appropriate markers (with multiple reverse X chromosomes) 2 days to 3 days after exposure to the test substance. The offspring of these females were used to evaluate the lethal effects of the test substance on mature sperm, late spermatocytes, early spermatocytes, spermatocytes and spermatogonia.
Each tube was mated with a treated male roller and a virgin fly according to the above procedure (i.e., one female fly was placed in each tube). Then, the F1 generation produced is mated with F1 to F2 at a female-female ratio of 1:1 or 1 = 2. After 12 days to 14 days, the F2 generation is observed and the number of lethality in the generation is recorded. If a tube of fruit flies contains an F generation female fly with a lethal factor in the parental X gene (i.e., no male mites with the above chromosome are found), then the female offspring with the same genotype female flies should be tested to confirm whether this lethality will reappear in the next generation.
The design of the experiment should have the predetermined sensitivity and measurement capacity. The number of fruit flies per distance should meet the needs of the experimental design. The frequency of white hair mutation is observed through the negative control group. When the mutation rate of the experimental group is close to that of the control group, the experiment should be repeated for verification. The experimental results should be confirmed in independent repeated experiments. 6 Experimental data and report
6.1 Results Summary
The test data should be tabulated to show the following information: the number of X chromosomes tested, the number of sterile males, and the number of lethal chromosomes produced in each dose group, each mating period, and each male individual exposed. The number of mutation clusters (clustcr) of different sizes in each male should also be included in the report. There are many statistical methods that can be used to evaluate sex-linked recessive lethal tests. The recessive lethal clusters produced by each sex individual should be considered and evaluated using appropriate statistical methods. Test data list: including the number of chromosomes tested, the number of sterile male flies, the number of lethal chromosomes for each exposure concentration, each male fly treated, and each mating period. Use statistical tests that are suitable for the experimental design. When evaluating the results, the biological significance and statistical significance of the test results should be considered at the same time. The observed results of the test group are significantly increased compared with the control group, and are judged as positive. When multiple doses are applied, the positive lethality shows a statistical dose-effect relationship; or at least one time point The positive lethality is repeatable and has a statistically significant increase, it should also be judged as positive.
6.2 Conclusions and Evaluation
There are several criteria for determining a positive response, one of which is a statistically significant dose-related increase in the frequency of recessive lethal mutations. Another criterion is a reproducible and statistically significant positive response at at least one test point. In this system, a test substance is considered non-mutagenic if it does not cause a statistically significant dose-related increase in the frequency of recessive lethal mutations and does not cause a reproducible and statistically significant positive response at any test point. Both biological and statistical significance should be considered in the evaluation. 6.3 Test Report
6.3.1 Strain: The population or strain of D. melanogaster used, age in days, number of male flies treated, number of sterile male flies, number of F2 culture tubes, number of F2 culture tubes without progeny, number of fruit flies tested, number of fruit flies carrying the lethal factor at each stage of germ cell development. The population or strain of Drosophila melanogaster used, age, number of male individuals treated, number of sterile male individuals, number of F2 generation cultures, number of F2 generation cultures without offspring, number of chromosomes inherited, number of chromosomes that died at each stage of germ cell development. 6.3.2 Test conditions: including detailed description of treatment and sampling scheme, dosage, toxicity data, appropriate negative (solvent) and positive controls.
6.3. 3 Criteria for judging lethal mutations. TTTKAONTKACA
GB/T27822—2011
6.3.4 Dose-effect relationship, if possible. 6.3.5 Statistical evaluation.
6.4 Interpretation of results
A positive result of the sex-linked recessive lethal test of Drosophila melanogaster indicates that the test sample causes mutations in the germ cells of Drosophila melanogaster under the test conditions; a negative result indicates that the test sample does not cause mutations in the germ cells of Drosophila melanogaster under the test conditions. TTTKANYKACA ||tt | et at.),Elsevier,Arnsterdam,1977:335-3735
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