Water, Air, & Soil Pollution

, 226:427 | Cite as

Harmful Effects of the Dermal Intake of Commercial Formulations Containing Chlorpyrifos, 2,4-D, and Glyphosate on the Common Toad Rhinella arenarum (Anura: Bufonidae)

  • Rafael C. Lajmanovich
  • Andrés M. Attademo
  • María F. Simoniello
  • Gisela L. Poletta
  • Celina M. Junges
  • Paola M. Peltzer
  • Paula Grenón
  • Mariana C. Cabagna-Zenklusen


Amphibians have complex life cycles with aquatic and terrestrial life and uncovered skins; therefore, they are exposed to chemical contamination, where dermal exposure is a significant route for pesticide uptake in both habitats. In this study, measurements in blood samples such as levels of butyrylcholinesterase (BChE), carboxylesterase (CbE), glutathione S-transferases (GST), thiobarbituric acid reactive substances (TBARS), modified alkaline comet assay (ACA) for detection of oxidized bases (FPG and Endo III sites), as well as the ratio of heterophils and lymphocytes (H/L), were evaluated as non-destructive biomarkers to monitor dermal pesticide exposure in male toads of Rhinella arenarum. Toads were exposed to a solution containing a nominal concentration of commercial formulations of the insecticide chlorpyrifos (CPF, 10 mg/L), and herbicides 2,4-D and glyphosate (GLY) (20 mg/L, respectively). After 48 h of exposure, the levels of plasma B-sterases (BChE and CbE) were inhibited (55 and 43 %, respectively) in toads exposed to CPF. Also, the activity of GST was inducted for dermal exposure to 2,4-D, as well as the levels of TBARS due to CPF exposure. Besides this, CPF and 2,4-D exposure induced oxidative DNA damage, and the H/L ratio decreased for the both herbicide exposures. Our results showed that exposure via dermal uptake to CPF, 2,4-D, and GLY in the common toad R. arenarum induced neurotoxicity, oxidative stress, and immunological depression. Thus, some blood biomarkers employed in our study (B-esterases, GST, levels of TBARS, ACA, and H/L ratio) might be used as predictors in health and ecological risk assessment of amphibian populations exposed to OP insecticides and herbicides.


Amphibians Dermal exposure Chlorpyrifos 2,4-D Glyphosate Blood non-destructive parameters 



This study was supported in part by the National Council for Scientific and Technical Research (CONICET), National Agency for Promotion of Science and Technology (ANPCyT), and the Course of Action for Research and Science Promotion (CAI + D-UNL). Authors are thankful to Dr. Andrew Collins (University of Oslo, Oslo, Norway) for providing the enzyme FPG and ENDO III. We also thank the anonymous reviewer for many valuable comments and suggestions.

Compliance with Ethical Standards

Animals used in this research have been treated according to ASIH and SSAR (2001) criteria and with approval from the Animal Ethics Committee of the Faculty of Biochemistry and Biological Sciences.

Conflict of Interest

The authors declare that they have no competing interests.


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Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • Rafael C. Lajmanovich
    • 1
    • 2
  • Andrés M. Attademo
    • 1
    • 2
  • María F. Simoniello
    • 2
  • Gisela L. Poletta
    • 1
    • 2
  • Celina M. Junges
    • 1
  • Paola M. Peltzer
    • 1
  • Paula Grenón
    • 2
  • Mariana C. Cabagna-Zenklusen
    • 2
  1. 1.National Council for Scientific and Technical Research (CONICET)Buenos AiresArgentina
  2. 2.Faculty of Biochemistry and Biological SciencesNational University of Littoral (FBCB-UNL)Santa FeArgentina

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