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Efficacy of essential oils from plants cultivated in the Amazonian Biome against gastrointestinal nematodes in sheep

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Abstract

The excessive use of anthelmintics to control nematodes has resulted in anthelminthic resistance. Essential oils (EOs) are a rich source of bioactive molecules that can be assessed for their ability to control resistant parasite populations. The aims of this study were to screen EOs from 10 plant species in vitro for anthelmintic activity against Haemonchus contortus, evaluate the cytotoxicity of those EOs in a human immortalized keratinocyte cell line (HaCaT), and test the most promising EO candidate in vivo in Santa Inês sheep. The efficacy was investigated in vitro using an egg hatch test (EHT) and a larval development test (LDT). EO cytotoxicity was evaluated with the sulforhodamine-B assay. In the in vivo experiment, 28 Santa Inês sheep naturally infected were distributed into groups: G1—Mentha arvensis (EO5), 200 mg kg−1; G2—menthol, 160 mg kg−1; G3—negative control; and G4—positive control (monepantel). EO5, from M. arvensis (86.7% menthol), had the lowest LC50 and LC90 values in the EHT (0.10, 0.27 mg mL−1, respectively), good performance in the LDT (0.015, 0.072 mg mL−1, respectively), and the lowest cytotoxicity (190.9 µg mL−1) in HaCaT cells. In the in vivo test, a single dose of the EO5 (200 mg kg−1 BW) had an efficacy of approximately 50% on days 1, 14, and 21; however, values were not significantly from day 0. Conversely, pure menthol at a dose of 160 mg kg−1 BW showed no in vivo efficacy. This can be attributed to key factors related to bioavailability and pharmacology of terpenes in the host organism, as well as to the fact that menthol is mainly excreted as glucuronides in urine. Thus, further studies should be conducted with formulation systems that deliver bioactives directly to the abomasum, focusing on terpenes, whose excretion route is mainly via faeces.

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Acknowledgements

We thank the Brazilian Agricultural Research Corporation (Embrapa, Project 03.11.01.023.00.00) and Amazonas Research Foundation (FAPEAM, Project 062.02821/2014) for funding. We also thank the National Council for Scientific and Technological Development (CNPq) for the scholarships granted to A. Figueiredo and F.A.S. Politi, and Dr. Christiane Pienna Soares of the Laboratory of Cytology and Cell Biology (UNESP), Araraquara (SP), for providing the HaCaT cells.

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Contributions

ACSC organized data, set statistical analyses and the manuscript final writing. AF performed the in vitro and in vivo trials. FGSP and IJM performed the cytotoxicity test. SNE provided and monitored the experimental animals. HRB and PEG performed chemical analyses and contributed to critical reading of the manuscript. FCMC contributed in collecting plant sample and extracting the essential oil.

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Correspondence to Ana Carolina S. Chagas.

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The authors declare that they have no competing interests.

Ethical approval

All experimental protocols were approved by the CPPSE Animal Care and Use Committee (No. 02/2014). Animals were under the care of a veterinary medical assistant during all 21 days of the experiment. The number of eggs per gram of faeces of the donor animal and the animals of the in vivo anthelmintic test was counted to monitor the level of infection during the experiment. After treatment, the animals were also observed for 1 h to monitor any toxicity symptoms caused by the essential oils, such as signs of discomfort, apathy, lethargy and/or drowsiness. None died or was in poor physical condition.

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Chagas, A.C.S., Figueiredo, A., Politi, F.A.S. et al. Efficacy of essential oils from plants cultivated in the Amazonian Biome against gastrointestinal nematodes in sheep. J Parasit Dis 42, 357–364 (2018). https://doi.org/10.1007/s12639-018-1007-x

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  • DOI: https://doi.org/10.1007/s12639-018-1007-x

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