Naslov
Cytotoxicity of herbicide atrazine residues from water on CHO (Chinese Hamster Ovary) cells in monolayer

Autori
Gaurina Srček, Višnja ; Kmetič, Ivana ; Kniewald, Zlatko ; Kniewald, Jasna

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Abstracts of the 3rd Croatian Scientific Conference about Biotechnology with International Participation - Biotechnology and Food, Zagreb, Croatia, February 17-20, 2003 / Kniewald, Zlatko - Zagreb : Hrvatsko Društvo za Biotehnologiju, 2003

Skup
3rd Croatian Scientific Conference about Biotechnology with International Participation - Biotechnology and Food

Mjesto i datum
Zagreb, Hrvatska, 17.02.2003. - 20.02.2003

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
atrazine residues; cytotoxicity; CHO-K1 cell line

Sažetak
The widespread use of agrochemicals as crop protection agents, wood preservatives, vector control agents or industrial/medicinal disinfectants, may result in exposure of man, animals and the environment to these chemicals. Humans may be exposed during production or application and to a lesser extent as consumers of food products and drinking water, which may contain trace levels of residues. Atrazine (2-chloro-4-ethylamino-6-isopropylamino-s-triazine) is a selective triazine herbicide, one of the most widely used pesticides to control broadleaf and grassy weeds in corn and other crops. Therefore the extensive use leads to its possible presence in ground water, as well as in raw and industrial food products. Even though atrazine is declared to be slightly toxic as toxicity class III, its possible ingestion via the food chain may present a risk for the reproductive process. By introducing in vitro methodology in toxicity testing, it is possible to evaluate the toxic influence of a chemical without using the great amount of experimental animals. In the present study, in order to determine cytotoxic effect of atrazine within the reproductive hormone-dependent tissues, the research was performed by using CHO K1 (Chinese Hamster Ovary) cell line in monolayer (CCL-61). Cell viability and number of cells were measured by Trypane Blue exclusion method in Fuchs-Rosenthal heamocytometer and lyzosomal activity measured by Neutral Red method. The cells were maintained at 37°C in an atmosphere of 95% air and 5% CO2 in Dulbecco medium supplemented with 10% fetal calf serum. The production of biomass was in T-bottle, and the cells were separated in the early logarithmic phase of growth. The initial concentrations of CHO cells were 2.5x10^4 cells/mL/well on 6-multiwell plates and after 72 h it succeeded 21x10^5 cells/mL in the control well. Cells were seeded 16 h before the treatment with atrazine in the concentrations of 5-80 microg/mL/well. The cytotoxicity was determined after 24, 48 and 72 h. The cell growth inhibition ranged after 72 h from 15 to 70% and was dose-responded. The IC_20, IC_50 and IC_80 were measured from the slope of % inhibition vs. log dose values. The values of IC_50 for atrazine after 72 h was 60.57 microg/mL with the both applied methods. Both colorimetric methods are acceptable for the quantitative determination of cytotoxic effects of atrazine on CHO K1 cell line. Application of cell lines from reproductive tissues like the Chinese hamster ovary enables determination of toxicity in the reproductive processes, as the first-step toxicity evaluation, by excluding the experiments with animals.

Izvorni jezik
Engleski

Znanstvena područja
Biotehnologija

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