Anti-protozoal activity of extracts from chicory (Cichorium intybus) against Cryptosporidium parvum in cell culture

Abstract

Cryptosporidium spp. are responsible for severe public health problems and livestock production losses. Treatment options are limited to only one drug available for human and bovine cryptosporidiosis, respectively, and both drugs exhibit only partial efficacy. Sesquiterpene lactones (SL) are plant bioactive compounds that function as a defence mechanism against herbivores. SL have demonstrated anti-parasitic properties against a range of parasitic taxa but knowledge about their anti-Cryptosporidium efficacy is limited. The effect of SL-rich leaf and root extracts from chicory (Cichorium intybus cv. Spadona) was investigated using human colon adenocarcinoma (HCT-8) cells infected with Cryptosporidium parvum. C. parvum oocysts were inoculated onto the cell monolayer and i) incubated for 4 hours with extracts (leaf and root extracts 300, 150, 75, 37.5, 18.75 and 9.375 μg/mL) in triplicates followed by incubation in bioactive free media (sporozoite invasion assays) or ii) incubated for 4 hours in bioactive free media followed by 48-hours incubation with extracts (growth inhibition assays). Extract toxicity on HCT-8 cells was assessed via water-soluble tetrazolium (WST)-1 assay prior to quantifying parasitic growth via immunofluorescence. Both extracts demonstrated dose-dependent inhibition in the growth inhibition assays (p = < 0.0001 for both extracts) but not in the invasion assays. Anti-parasitic activity did not appear to be solely related to SL content, with the extract with lower SL content (leaf) exhibiting higher inhibition at 300 μg/ml. However, given the limited treatment options available for Cryptosporidium spp., our study encourages further investigation into the use of chicory extracts to identify novel active compound(s) inhibiting these protozoa.

The Common Cichory (Cichorium intybus L.) as a Source of Extracts with Health-Promoting Properties—A Review

“…Woolsey et al demonstrated the efficacy of chicory leaf and root extracts against Cryptosporidium parvum , with the lower SL extract (leaf) showing greater inhibition. Based on the results, the authors suggest that the antiparasitic activity does not appear to be related only to the SL content [ 45 ]. Marley et al conducted a study on lambs that acquired parasites by natural transmission.…”

Activity of (1-benzyl-4-triazolyl)-indole-2-carboxamides against Toxoplasma gondii and Cryptosporidium parvum

“…As such, we allowed 2 h for the establishment of infection in this study. Furthermore, the 2–4 h time point has been used by many research groups (including ours) in the recent past to assess the in vitro effect of compounds on these parasites before and after invasion of host cells ( Jumani et al, 2019 ; Woolsey et al, 2019 ; Funkhouser-Jones et al, 2020 ; Li et al, 2020 ; Ma et al, 2020 ; Murakoshi et al, 2020 ). When the cultures were analyzed at 72 h post-infection, five compounds (JS-2-22, 27, 28, 30 and 44) were found to have concentration-dependent activity of reducing C. parvum growth and proliferation, regardless of whether treatment started immediately or 2 h after infection of host cells ( Fig.…”

Contemporary Insights on Antibacterial and Pharmacological Importance of Cichorium intybus: A Compendious Review

“…As a hepatoprotective agent, it also decreases the threat of liver cirrhosis [6]. Other pharmacological application of chicory includes antihypoglycemic, antihyperlipidemic [90,91], anticancer [92], anti-allergic [93], anti-ulcerogenic [94], gastroprotective [95], anti-protozoal [96], and wound healing activities [23]. It has evaluated to be an anti-hyperuricemia agent [97], and also depicted anti-rheumatic effects [98].…”