Lethal and Sublethal Effects of the Herbicide Atrazine in the Early Stages of Development of Physalaemus gracilis (Anura: Leptodactylidae)

  • Camila F. Rutkoski
  • Natani Macagnan
  • Cassiane Kolcenti
  • Guilherme V. Vanzetto
  • Paola F. Sturza
  • Paulo A. Hartmann
  • Marilia T. Hartmann


Water sources used as reproductive sites by crying frog, Physalaemus gracilis, are extensively associated with agroecosystems in which the herbicide atrazine is employed. To evaluate the lethal and sublethal effects of atrazine commercial formulation, acute and chronic toxicity tests were performed in the embryonic phase and the beginning of the larval phase of P. gracilis. Tests were started on stage 19 of Gosner (Herpetologica 16:183–190, 1960) and performed in 24-well cell culture plates. Acute tests had a duration of 96 h with embryo mortality monitoring every 24 h. Chronic assays contemplated the transition from the embryonic to larval stages and lasted 168 h. Every 24 h the embryos/larvae were observed for mortality, mobility, and malformations. The LC50 of atrazine determined for P. gracilis embryos was 229.34 mg L−1. The sublethal concentrations did not affect the development of the larvae but were observed effects on mobility and malformations, such as spasmodic contractions, reduced mobility, malformations in mouth and intestine, and edema arising. From 1 mg L−1 atrazine, the exposed larvae began to have changes in mobility and malformations. The atrazine commercial formulation has caused early life effects of P. gracilis that may compromise the survival of this species but at higher concentrations than recorded in the environment, so P. gracilis can be considered tolerant to this herbicide at environmentally relevant concentrations.



Valuable help in laboratorial work was provided by Jessica Herek, Jéssica G. Slaviero, and Gregori B. Bieniek. The authors are grateful to the Federal University of Fronteira Sul—UFFS for providing logistical support. Camila Rutkoski, Natani Macagnan, and Cassiane Kolcenti were supported by fellowship from Fundação de Amparo a Pesquisa do Estado do Rio Grande do Sul—FAPERGS.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This study was licensed by IBAMA (50398-1) and authorized by the Ethics Committee for Animal Use of the Federal University of Fronteira Sul.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Camila F. Rutkoski
    • 1
  • Natani Macagnan
    • 1
  • Cassiane Kolcenti
    • 1
  • Guilherme V. Vanzetto
    • 1
  • Paola F. Sturza
    • 1
  • Paulo A. Hartmann
    • 1
  • Marilia T. Hartmann
    • 1
  1. 1.Ecology and Conservation LaboratoryFederal University of Fronteira SulErechimBrazil

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