In vitro anthelmintic effects of Spigelia anthelmia protein fractions against Haemonchus contortus

Abstract

Gastrointestinal nematodes are a significant concern for animal health and well-being, and anthelmintic treatment is mainly performed through the use of chemical products. However, bioactive compounds produced by plants have shown promise for development as novel anthelmintics. The aim of this study is to assess the anthelmintic activity of protein fractions from Spigelia anthelmia on the gastrointestinal nematode Haemonchus contortus. Plant parts were separated into leaves, stems and roots, washed with distilled water, freeze-dried and ground into a fine powder. Protein extraction was performed with sodium phosphate buffer (75 mM, pH 7.0). The extract was fractionated using ammonium sulfate (0–90%) and extensively dialyzed. The resulting fractions were named LPF (leaf protein fraction), SPF (stem protein fraction) and RPF (root protein fraction), and the protein contents and activities of the fractions were analyzed. H. contortus egg hatching (EHA), larval exsheathment inhibition (LEIA) and larval migration inhibition (LMIA) assays were performed. Proteomic analysis was conducted, and high-performance liquid chromatography (HPLC) chromatographic profiles of the fractions were established to identify proteins and possible secondary metabolites. S. anthelmia fractions inhibited H. contortus egg hatching, with LPF having the most potent effects (EC50 0.17 mg mL-1). During LEIA, SPF presented greater efficiency than the other fractions (EC50 0.25 mg mL-1). According to LMIA, the fractions from roots, stems and leaves also reduced the number of larvae, with EC50 values of 0.11, 0.14 and 0.21 mg mL-1, respectively. Protein analysis indicated the presence of plant defense proteins in the S. anthelmia fractions, including protease, protease inhibitor, chitinase and others. Conversely, secondary metabolites were absent in the S. anthemia fractions. These results suggest that S. anthelmia proteins are promising for the control of the gastrointestinal nematode H. contortus.

Myracrodruon urundeuva seed exudates proteome and anthelmintic activity against Haemonchus contortus

“…Diverse proteins have been studied for nematicidal activity [ 3 , 44 , 45 ]. The proteomic profiles of SEX, SEXF1, and SEXF2 revealed the presence of proteins that potentially can affect the nematode development cycle.…”

“…For instance, proteins from Spigelia anthelmia inhibited egg hatching, larval exsheathment, and larval migration of H . contortus [ 3 ]. Cysteine proteinases from Carica papaya displayed anthelmintic activity against Protospirura muricola [ 4 ] and Trichuris trichiura , as revealed by randomized controlled trials conducted in pigs [ 5 ].…”

The Effect of Season Upon the Life Cycle of Haemonchus Contortus in Experimentally Infected Lambs

“…After rinsing, the larvae are subjected to a sodium hypochlorite solution (2% w/v) and exsheathment is analyzed under a microscope every 10 minutes for a period of one hour (Bahuaud et al, 2006). Since its initial development, the larval exsheathment bleach assay has been used in many studies that focus on testing various tanniferous plant species for anthelmintic properties (Alonso-Diaz et al, 2008;Araújo et al, 2017).…”

“…These life cycle stages are typically examined using experimentally infected sheep and goats on a year-round basis to provide fecal eggs and larvae used in in-vitro and invivo assays (Brunet et al, 2007;Alonso-Diaz et al, 2008;Araújo et al, 2017;Assis et al 2003;Barone et al, 2018).…”

“…Multiple assays have been developed to assess exsheathment in vitro (Bahuaud et al, 2006;Alonso-Diaz et al, 2008;Araújo et al, 2017;Conder and Johnson, 1996;Barone et al 2018). The sodium hypochlorite method (Bahuaud et al, 2006) is more commonly utilized (Alonso-Diaz et al, 2008;Araújo et al, 2017), however it has been suggested that the CO2 method improves viability and infectivity (Conder and Johnson, 1996). A considerable amount of variability has been observed across in vivo studies in relation to methodology, as well as time for exsheathment to occur Hertzberg et al, 2002;Brunet et al, 2007).…”