Cultural, morphological and biochemical properties of Salmonella typhimurium indicator bacteria used in the Ames test and their applicability for the assessment of the test system quality

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Introduction. The Ames test has been widely used to study the mutagenicity of chemicals. In view of the implementation of a quality management system in the test facilities, the cultural, morphological and biochemical properties of strains used in the Salmonella/microsome test system were studied. An assessment of their applicability as additional characteristics of quality of the tester strain was made.

Materials and methods. The study of cultural and morphological characteristics of S. typhimurium strains was carried out using the commercial media. The biochemical properties of S. typhimurium cultures were evaluated using a Vitek-2 system.

Results. Studies have shown that the procedure of marker verification for confirming the reference properties of tester strains, in addition to evaluation of the spontaneous mutation level and phenotypic characteristics (-his phenotype, the presence or absence of the R-factor plasmids, rfa- and ∆uvrB mutations), can be extended by periodically assessing their biochemical properties. Most of the biochemical characteristics of bacteria of the Salmonella typhimurium serovar are inherent for tester strains. Compared to the wild-type bacteria аtypical features of tester strains are a negative reaction in the hydrogen sulfide production test, positive activity of tyrosine arylamidase, the ability to utilize 5-keto-D-gluconate, unstable activity of alpha-galactosidase. Differential selective media used to maintain wild-type Salmonella cultures are not universal for the growth of strains of the Salmonella/microsome test system. When choosing media for cytotoxicity, survival, or sterility assessment, it is necessary to take into consideration the growth characteristics of the Salmonella tester strains on different media: the ability to grow on selective media decreases in the series of Endo-agar > Salmonella Shigella -agar > Bismuth-sulfite agar > Ploskirev’s agar.

Limitations. The research is limited to the study of cultural, morphological and biochemical characteristics of S. typhimurium, but not Escherichia coli.

Conclusion. The described cultural and morphological properties of tested strains on different media can be used to assess cytotoxicity/survival of treated cultures performing studies according to OECD 471. The use of additional biochemical markers of authenticity and morphological properties of tester strains will help to ensure the quality of studies using the Ames test in the processes of conservation, reproduction, and routine testing.

Compliance with ethical standards. The study does not need the approval of the biomedical ethics committee or other documents.

Contribution:
Egorova O.V. — the concept and design of the study, the collection and processing of material, writing the text;
Mikheev P.V. — collection and processing of material;
Ilyushina N.A. — analysis of materials, writing the text.
All authors are responsible for the integrity of all parts of the manuscript and approval of the manuscript final version

Conflict of interest. The authors declare no conflict of interest.

Acknowledgement. The study had no sponsorship.

Received: March 23, 2023 / Accepted: June 7, 2023 / Published: July 30, 2023

作者简介

Olga Egorova

Federal Scientific Center of Hygiene named after F.F. Erisman of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

编辑信件的主要联系方式.
Email: egorovaov@fferisman.ru
ORCID iD: 0000-0003-4748-8771

MD, PhD, senior researcher of the department of genetic toxicology, Federal Scientific Center of Hygiene named after F.F. Erisman of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Mytishchi, 141014, Russian Federation.

e-mail: egorovaov@fferisman.ru

俄罗斯联邦

Pavel Mikheev

Federal Scientific Center of Hygiene named after F.F. Erisman of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Email: noemail@neicon.ru
ORCID iD: 0000-0002-3376-0677
俄罗斯联邦

Galina Trukhina

Federal Scientific Center of Hygiene named after F.F. Erisman of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Email: noemail@neicon.ru
ORCID iD: 0000-0001-9955-7447
俄罗斯联邦

Nataliya Ilyushina

Federal Scientific Center of Hygiene named after F.F. Erisman of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Email: noemail@neicon.ru
ORCID iD: 0000-0001-9122-9465
俄罗斯联邦

参考

  1. Chugunova E.O., Tatarnikova N.A., Prokhorova T.S. Development of an Accelerated Method and Construction of a Nutrient Medium for the Isolation of Salmonella from Foods [Razrabotka uskorennogo sposoba i konstruirovanie pitatel’noy sredy dlya vydeleniya sal’monell iz pishchevykh produktov]. Perm’: Prokrost”; 2017. (in Russian)
  2. Ames B.N. The detection of chemical mutagens with enteric bacteria. In: Hollaender A., ed. Chemical Mutagens. Boston: Springer; 1971: 267–82. https://doi.org/10.1007/978-1-4615-8966-2_9
  3. Zeiger E. The test that changed the world: The Ames test and the regulation of chemicals. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2019; 841: 43–8. https://doi.org/10.1016/j.mrgentox.2019.05.007
  4. OECD Library. Test 471: 2020, IDT. Bacterial reverse mutation test. Available at: https://www.oecd-ilibrary.org/environment/test-no-471-bacterial-reverse-mutation-test_9789264071247-en
  5. Maron D.M., Ames B.N. Revised methods for the Salmonella mutagenicity test. Mutat. Res. 1983; 113(3-4): 173–215. https://doi.org/10.1016/0165-1161(83)90010-9
  6. Vijay U., Gupta S., Mathur P., Suravajhala P., Bhatnagar P. Microbial mutagenicity assay: Ames test. Bio. Protoc. 2018; 8(6): e2763. https://doi.org/10.21769/BioProtoc.2763
  7. Zeiger E. Bacterial mutation assays. Methods Mol. Biol. 2013; 1044: 3–26. https://doi.org/10.1007/978-1-62703-529-3_1
  8. Escobar P.A., Kemper R.A., Tarca J., Nicolette J., Kenyon M., Glowienke S., et al. Bacterial mutagenicity screening in the pharmaceutical industry. Mutat. Res. 2013; 752(2): 99–118. https://doi.org/10.1016/j.mrrev.2012.12.002
  9. Mortelmans K. A perspective on the development of the Ames Salmonella/mammalian-microsome mutagenicity assay. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2019; 841: 14–6. https://doi.org/10.1016/j.mrgentox.2019.04.004
  10. Schoeny R., Cross K.P., DeMarini D.M., Elespuru R., Hakura A., Levy D.D., et al. Revisiting the bacterial mutagenicity assays: Report by a workgroup of the International Workshops on Genotoxicity Testing (IWGT). Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2020; 849: 503137. https://doi.org/10.1016/j.mrgentox.2020.503137
  11. Egorova O.V. To the procedure of confirmation of the laboratory’s competence in performing mutagenicity assessment using the Ames test. Toksikologicheskiy vestnik. 2021; 29(4): 4–13. https://doi.org/10.36946/0869-7922-2021-29-4-4-13 https://elibrary.ru/dpghdx (in Russian)
  12. Levy D.D., Hakura A., Elespuru R.K., Escobar P.A., Kato M., Lott J., et al. Demonstrating laboratory proficiency in bacterial mutagenicity assays for regulatory submission. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2019; 848: 403075. https://doi.org/10.1016/j.mrgentox.2019.07.005
  13. Hamel A., Roy M., Proudlock R. The bacterial reverse mutation test. Chapter 4. In: Proudlock R., ed. Genetic Toxicology Testing. A Laboratory Manual. Academic Press; 2016: 79–138.
  14. Mortelmans K., Zeiger E. The Ames Salmonella/microsome mutagenicity assay. Mutat. Res. 2000; 455(1–2): 29–60. https://doi.org/10.1016/s0027-5107(00)00064-6
  15. Andrusevich A.S., Tyapsha Yu.I., Dubanevich O.V., Strel’chenya I.I. Biochemical properties of museum strains Salmonella Dublin, Salmonella cholerae suis, Salmonella typhimurium. Epizootologiya, immunobiologiya, farmakologiya i sanitariya. 2020; (1): 39–44. https://elibrary.ru/gxlrwd (in Russian)
  16. Busch D.B., Archer J., Amos E.A., Hatcher J.F., Bryan G.T. A protocol for the combined biochemical and serological identification of the Ames mutagen tester strains as Salmonella typhimurium. Environ. Mutagen. 1986; 8(5): 741–51. https://doi.org/10.1002/em.2860080509
  17. Speck W.T., Ellner P.D., Rosenkranz H.S. Mutagenicity testing with Salmonella typhimurium strains. I. Unusual phenotypes of the tester strains. Mutat. Res. 1975; 28(1): 27–30. https://doi.org/10.1016/0027-5107(75)90310-3
  18. Pavlova I.B., Zuev V.S. Biochemical properties of Salmonella living in the aquatic environment. Veterinarnaya patologiya. 2007; (1): 81–5. (in Russian)
  19. Tan C.S., Pasculescu A., Lim W.A., Pawson T., Bader G.D., Linding R. Positive selection of tyrosine loss in metazoan evolution. Science. 2009; 325(5948): 1686–8. https://doi.org/10.1126/science.1174301
  20. Begüm Ç., Doğruöz G.N. The biotechnological potentials of bacteria isolated from Parsık cave, Turkey: Measuring the enzyme profiles, antibiotic resistance and antimicrobial activity in bacteria. Johnson Matthey Technol. Rev. 2020; 64(4): 396–406. https://doi.org/10.1595/205651320X15923194903811
  21. Bausch C., Peekhaus N., Utz C., Blais T., Murray E., Lowary T., Conway T. Sequence analysis of the GntII (subsidiary) system for gluconate metabolism reveals a novel pathway for L-idonic acid catabolism in Escherichia coli. J Bacteriol. 1998; 180(14): 3704–10. https://doi.org/10.1128/JB.180.14.3704-3710.1998
  22. Irai A.S., Ezawa K.U., Wamura M.S., Tsushima T.M., Imura T.S. Effect of glucose on the mutagenic sensitivity of Salmonella typhimurium tester strains. Mutat. Res. Environ. Mutagen. Relat. Subj. 1978; 54(2): 253–4.
  23. Porwollik S., Wong R.M., Sims S.H., Schaaper R.M., DeMarini D.M., McClelland M. The ∆uvrB mutations in the Ames strains of Salmonella span 15 to 119 genes. Mutat. Res. 2001; 483(1–2): 1–11. https://doi.org/10.1016/s0027-5107(01)00239-1
  24. Matsumura S., Ito Y., Morita O., Honda H. Genome resequencing analysis of Salmonella typhimurium LT-2 strains TA98 and TA100 for the establishment of a next-generation sequencing-based mutagenicity assay. J. Appl. Toxicol. 2017; 37(9): 1125–8. https://doi.org/10.1002/jat.3463

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