Skip to main content
SpringerLink
Log in

Genotoxic effects of commercial formulations of Chlorpyrifos and Tebuconazole on green algae

  • Published:
Ecotoxicology Aims and scope Submit manuscript

Abstract

The alkaline single-cell gel electrophoresis assay (comet assay) was used for the study of the genotoxic effects of insecticide Chlorpyrifos and fungicide Tebuconazole (commercial formulations) on two freshwater green algae species, Pseudokirchneriella subcapitata and Nannocloris oculata, after 24 h of exposure. The percentage of DNA in tail of migrating nucleoids was taken as an endpoint of DNA impairment. Cell viability was measured by fluorometric detection of chlorophyll “a” in vivo and the determination of cell auto-fluorescence. Only the higher concentration of Chlorpyrifos tested resulted to affect significantly the cell viability of P. subcapitata, whereas cells of N. oculata were not affected. Tebuconazole assayed concentrations (3 and 6 mg/l) did not affect cell viability of both species. The results of comet assay on P. subcapitata showed that Chlorpyrifos concentration evaluated (0.8 mg/l) exerted a genotoxic effects; while for the other specie a concentration of 10 mg/l was needed. Tebuconazole was genotoxic at 3 and 6 mg/l for both species. The comet assay evidenced damage at the level of DNA simple strains molecule at pesticide concentrations were cytotoxicity was not evident, demonstrating that algae are models to take into account in ecological risk assessments for aquatic environments.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Akcha F, Arzul G, Rousseau R, Bardouil M (2008) Comet assay in phytoplankton as biomarker of genotoxic effects of environmental pollution. Mar Environ Res 66:59–61

    Article  CAS  Google Scholar 

  • Ali D, Nagpure NS, Kumar S, Kumar R, Kushwaha B (2008) Genotoxicity assessment of acute exposure of chlorpyrifos to freshwater fish Channa punctatus (Bloch) using micronucleus assay and alkaline single-cell gel electrophoresis. Chemosphere 71:1823–1831

    Article  CAS  Google Scholar 

  • Aoyama K, Iwahori K, Miyata N (2003) Application of Euglena gracilis cells to comet assay: evaluation of DNA damage and repair. Mutat Res 538:155–162

    Article  CAS  Google Scholar 

  • Boyd-Boland AA, Magdic S, Pawliszyn JB (1996) Simultaneous determination of 60 pesticides in water using solid-phase microextraction and gas chromatography–mass spectrometry. Analyst 121:929–937

    Article  CAS  Google Scholar 

  • CASAFE (Cámara Argentina de Productos Sanitarios y Fertilizantes). 2013. Registro de plaguicidas. Informe de relevamiento de uso de plaguicidas a nivel nacional. http://www.casafe.org/.2013. Accessed May 2013

  • Ciniglia C, Cascone C, Lo Giudice R, Pinto G, Pollio A (2005) Application of methods for assessing the geno- and cytotoxicity of Triclosan to C. ehrenbergii. J Hazard Mater 122:227–232

    Article  CAS  Google Scholar 

  • Cotelle S, Férard JF (1999) Comet assay in genetic ecotoxicology: a review. Environ Mol Mutagen 34:246–255

    Article  CAS  Google Scholar 

  • Debenest T, Silvestre J, Coste M, Delmas F, Pinelli E (2008) Herbicide effects on freshwater benthic diatoms: induction of nucleus alterations and silica cell wall abnormalities. Aquat Toxicol 88:88–94

    Article  CAS  Google Scholar 

  • Desai SR, Verlecar XN, Goswami U (2006) Genotoxicity of cadmium in marine diatom Chaetoceros tenuissimus using the alkaline Comet assay. Ecotoxicology 15:359–363

    Article  CAS  Google Scholar 

  • Dhawan A, Bajpayee M, Parmar D (2008) Comet assay: a reliable tool for the assessment of DNA damage in different models. Cell Biol Toxicol 25:5–32

    Article  Google Scholar 

  • Di Marzio WD, Sáenz ME, Lemiere S, Vasseur P (2005) Improved single-cell gel electrophoresis assay for detecting DNA damage in Eisenia foetida. Environ Mol Mutagen 46:246–252

    Article  Google Scholar 

  • Erbes M, Weßler A, Obst U, Wild A (1997) Detection of primary DNA damage in Chlamydomonas reinhardtii by means of modified microgel electrophoresis. Environ Mol Mutagen 30:448–458

    Article  CAS  Google Scholar 

  • Fucic A, Markovi D, Herceg Z, Gamulin M, Katic J, Stojkovic R, Ferencic Z, Mildner B, Jazbec AM, Dobranic T (2008) Developmental and transplacental genotoxicology: fluconazole. Mutat Res 657:43–47

    Article  CAS  Google Scholar 

  • Goetz AK, Dix DJ (2009) Mode of action for reproductive and hepatic toxicity inferred from a genomic study of triazole antifungals. Toxicol Sci 110:449–462

    Article  CAS  Google Scholar 

  • Hornsby AG, Wauchope R, Herner AE (1996) Pesticide properties in the environment. Springer, New York, p 227

    Book  Google Scholar 

  • Jha A (2008) Ecotoxicological applications and significance of the comet assay. Mutagenesis 23:207–221

    Article  CAS  Google Scholar 

  • Kumar N, Bora A, Kumar R, Ambi MK (2012) Differential effects of agricultural pesticides endosulfan and tebuconazole on photosynthetic pigments, metabolism and assimilating enzymes of three heterotrophic, filamentous Cyanobacteria. J Biol Environ Sci 6:67–75

    Google Scholar 

  • Kumarravel TS, Jha AN (2006) Reliable Comet assay measurements for detecting DNA damage induced by ionising radiation and chemicals. Mutat Res 605:7–16

    Article  Google Scholar 

  • Lee RF, Steinert S (2003) Use of the single cell gel electrophoresis/comet assay for detecting DNA damage in aquatic (marine and freshwater) animals. Mutat Res 544:43–64

    Article  CAS  Google Scholar 

  • Martinez RS (2013) Evaluación de los efectos sobre la tasa de división celular y genotóxicos de formulaciones comerciales de plaguicidas sobre dos especies de algas verdes (Chlorophyceae) dulceacuícolas. Graduation Thesis, Universidad Nacional de Luján, Luján

  • Mehta A, Verma RS, Srivastava N (2008) Chlorpyrifos-induced DNA damage in rat liver and brain. Environ Mol Mutatgen 49:426–433

    Article  CAS  Google Scholar 

  • Nesnow S, Padgett WT, Moore T (2011) Propiconazole induces alterations in the hepatic metabolome of mice: relevance to propiconazole-induced hepatocarcinogenesis. Toxicol Sci 120:297–309

    Article  CAS  Google Scholar 

  • Office of Environmental Health Hazard Assessment (OEHHA) (2011) Triazole Antifungal Agents. WHO, Geneve

    Google Scholar 

  • Prado R, García R, Rioboo G, Herrero C, Abalde J, Cid A (2009) Comparison of the sensitivity of different toxicity test endpoints in a microalga exposed to the herbicide paraquat. Environ Internat 35:240–247

    Article  CAS  Google Scholar 

  • Qiao D, Seidler FM, Violin J, Slotkin T (2003) Nicotine is a developmental neurotoxicant and neuroprotectant: stage-selective inhibition of DNA synthesis coincident with shielding from effects of chlorpyrifos. Develop Brain Res 147:183–190

    Article  CAS  Google Scholar 

  • Rahman MM, Mahboob K, Danadevi B, Banu B, Grover P (2002) Assessment of genotoxic effects of chloropyrifos and acephate by the comet assay in mice leucocytes. Mutat Res 516:139–147

    Article  CAS  Google Scholar 

  • Sáenz ME (2000) Estudios de los efectos de contaminantes sobre poblaciones algales de agua dulce. Doctoral Thesis, Facultad de Ciencias Naturales y Museo, Universidad Nacional de La Plata, La Plata

  • Sáenz ME, Di Marzio WD, Ambrini G (2009) Toxicidad de insecticidas organofosforados utilizados en cultivo de soja sobre productores primarios. Rev Arg Ecotoxicol Contam Amb 1:29–36

    Google Scholar 

  • Sáenz ME, Di Marzio WD, Alberdi JL (2012a) Assessment of Cyfluthrin commercial formulation on growth, photosynthesis and catalase activity of green algae. Pest Biochem Physiol 104:50–57

    Article  Google Scholar 

  • Sáenz ME, Di Marzio WD, Alberdi JL (2012b) Effects of a commercial formulation of Cypermethrin used in Biotech Soybean Crops on Growth and Antioxidant Enzymes of Freshwater Algae. Inter J Environ Prot 2:15–22

    Google Scholar 

  • Sastre MP, Vernet M, Steinert S (2001) Single-cell gel/comet assay applied to the analysis of UV radiation–induced DNA damage in Rhodomonas sp. (Cryptophyta). Photochem Photobiol 74:55–60

    Article  CAS  Google Scholar 

  • Schubert LE (2003) Nonmotile coccoid and colonial green algae. In: Wehr JD, Sheath RG (eds) Freshwater Algae of North America. Academic Press, New York

    Google Scholar 

  • Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H, Miyamae Y, Rojas E, Ryu JC, Sasaki YF (2000) Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutag 35:206–221

    Article  CAS  Google Scholar 

  • Torres MA, Barros MP, Campos SC, Pinto E, Rajamani S, Sayre RT, Colepicolo P (2008) Biochemical biomarkers in algae and marine pollution: a review. Ecotoxicol Environ Saf 71:1–15

    Article  CAS  Google Scholar 

  • Uhl M, Plewa MJ, Majer BJ, Knasmüller S (2003) Basic principles of genetic toxicology with an emphasis on plant bioassays. In: Maluszynska J, Plewa M (eds) Bioassays in Plant Cells for Improvement of Ecosystem and human health: A course manual. Wydawnictvo Uniwersytetu Ślaskiego, Katowice, Poland, pp 11–30

  • US Environmental Protection Agency, EPA (2002a) Selenatrum capricornutum growth test. In: short-term methods for estimating the chronic toxicity of effluents and receiving water to freshwater organisms. EPA/821-R-02-013 U.S. Environmental Monitoring and Support Laboratory Office of Research and Development, Cincinnati, Ohio

  • US Environmental Protection Agency, EPA (2002b) Office of Pesticide Programs List of Chemicals Evaluated for Carcinogenic Potential

  • US Environmental Protection Agency EPA (2010) Tebuconazole; Pesticide tolerances Federal Register.752442124428

  • US Environmental Protection Agency, EPA (2012) Estimation Programs Interface Suite™ for Microsoft® Windows, v 4.11. United States Environmental Protection Agency, Washington, DC, USA

  • Vasseur P, Cossu-Leguille C (2003) Biomarkers and community indices as complementary tools for environmental safety. Environ Intern 28:711–717

    Article  CAS  Google Scholar 

  • Villem A (2011) Algae Pseudokirchneriella subcapitata in environmental hazard evaluation of chemicals and synthetic nanoparticles. PhD Thesis, Institute of Agricultural and Environmental Sciences, Estonian University of Life Sciences

  • Vindas R, Ortiz F, Ramírez V, Cuenca P (2004) Genotoxicidad de tres plaguicidas utilizados en la actividad bananera de Costa Rica. Rev Biol Trop 52:601–609

    Google Scholar 

  • Yaqin K (2006) Ecotoxicological Assessment of Aquatic Genotoxicity Using the Comet Assay. Hayati J Bios 13:124–130

    Google Scholar 

  • Zambonin CG, Cilenti A, Palmisano F (2002) Solid-phase microextraction and gas chromatography-mass spectrometry for the rapid screening of triazole residues in wine and strawberries. J Chromatogr 967:255–260

    Article  CAS  Google Scholar 

  • Zar JH (2010) Biostatistical Analysis. Pearson, Boston

    Google Scholar 

Download references

Acknowledgments

This research was supported by Department of Basic Sciences and Secretary of Sciences and Technique founds from 3.5, National University of Luján, Buenos Aires, Argentina.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Ecotoxicology Research Program, Department of Basic Sciences, National University of Luján, P.O. Box 221, 6700, Luján, Buenos Aires, Argentina

    Ricardo Santiago Martinez, Walter Darío Di Marzio & María Elena Sáenz

  2. National Council of Scientific and Technical Researches CONICET, Av. Rivadavia 1917, 1023, Buenos Aires, Argentina

    Ricardo Santiago Martinez, Walter Darío Di Marzio & María Elena Sáenz

Authors
  1. Ricardo Santiago Martinez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Walter Darío Di Marzio
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. María Elena Sáenz
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to María Elena Sáenz.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Martinez, R.S., Di Marzio, W.D. & Sáenz, M.E. Genotoxic effects of commercial formulations of Chlorpyrifos and Tebuconazole on green algae. Ecotoxicology 24, 45–54 (2015). https://doi.org/10.1007/s10646-014-1353-0

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10646-014-1353-0

Keywords

Search

Navigation