In Vitro Nematocidal Effect and Anthelmintic Activity of Artemisia cina Against Haemonchus contortus in Gerbils and Relative Expression of Hc29 Gene in Transitional Larvae (L3-L4)

Acta Parasitol. 2021 Sep;66(3):938-946. doi: 10.1007/s11686-021-00364-w. Epub 2021 Mar 15.

Abstract

Purpose: (1) To assess the in vitro activity of Artemisia cina against Haemonchus contortus L3 (HcL3) and in transitional (L3-L4) larvae (HcTrL3-L4); (2) to quantify the relative expression of the Hc29 gene in HcTrL3-L4 exposed to the A. cina n-hexane extract; and (3) to assess the anthelmintic activity (AA) of the A. cina organic extracts in gerbils artificially infected with H. contortus (HcArt/inf/gerbs).

Methods: The in vitro assay was carried out in 96-well microtitration plates. The following A. cina extracts: ethyl acetate (Ac-EtOAcEx), n-hexane (Ac-n-HexEx), and methanol (Ac-MethEx) were assessed at 1 and 2 mg/mL against HcL3 and HcTrL3-L4 at 24 h exposure. Relative expression of the Hc29 gene in HcTrL3-L4 was obtained by RT-PCR. For assessing the AA, six groups of five HcArt/inf/gerbs were used. Groups were treated orally with 4 mg/kg BW of A. cina extracts. Five days after treatment, the gerbils were necropsied and nematodes counted.

Results: The highest in vitro activities (75 and 82.6%) were shown by Ac-n-HexEx at 1 and 2 mg/mL, respectively. For HcTrL3-L4 the highest in vitro activities (69 and 23%) were shown by Ac-n-HexEx and isoguaiacine at 0.625 mg/mL, respectively. Also, upregulation of H. contortus Hc29 gene by 13- and 80-fold (p < 0.01) was observed on the HcTrL3-L4 stage after exposure to Ac-n-HexEx extract and isoguaiacine at 0.078 mg/mL, respectively. Reduction percentage was 100% in HcArt/inf/gerbs treated with Ac-n-HexEx.

Conclusions: We conclude that the Ac-n-HexEx and isoguaiacine compound had anthelmintic efficacy against H. contortus and L3 and HcTrL3-L4.

Keywords: Artemisia cina; Artemisinin; Haemonchus contortus; Isoaguaicine; Meriones unguiculatus.

MeSH terms

  • Animals
  • Anthelmintics* / pharmacology
  • Anthelmintics* / therapeutic use
  • Antinematodal Agents / pharmacology
  • Antinematodal Agents / therapeutic use
  • Artemisia*
  • Gerbillinae
  • Haemonchiasis* / drug therapy
  • Haemonchiasis* / veterinary
  • Haemonchus* / genetics
  • Larva
  • Plant Extracts / pharmacology

Substances

  • Anthelmintics
  • Antinematodal Agents
  • Plant Extracts