Evaluation of Antioxidant and Anthelmintic Properties of Tithonia diversifolia Extracts Against Gastrointestinal Nematode Eggs Using In Vitro Assays

Summary

This study evaluated the biological activities of Tithonia diversifolia extracts using radical-based methods and an in vitro egg hatching assay (EHA) to assess their effects on gastrointestinal nematodes in small ruminants. The methodology and key findings are described, highlighting the potential of these extracts for controlling parasites.

Abstract

Parasitic diseases caused by gastrointestinal nematodes (GIN) are considered important limiting factors for livestock production worldwide, as they significantly compromise animal welfare and increase production costs. For years, parasite control has relied exclusively on the use of synthetic drugs. Resistance of GIN to commercial anthelmintics has been increasingly reported throughout the world. Therefore, alternative approaches have emerged for controlling GIN in farm animals, including the exploration of plants with bioactive potential. Tithonia diversifolia can be harvested year-round, and all parts of the plant have been used in folk medicine for a wide range of diseases. This study evaluated the in vitro antioxidant properties of water and 70% aqueous acetone extracts of T. diversifolia aerial parts, including leaves, branches, and flowers, using radical-based methods, and assessed its anthelmintic potential through a parasite model, employing the egg hatching assay (EHA) with water extracts. Additionally, the total polyphenolic content of the extracts was determined. These findings highlight T. diversifolia as a promising natural source of antioxidant and anthelmintic compounds.

Introduction

Parasitic diseases caused by gastrointestinal nematodes (GIN) are considered important limiting factors for livestock production worldwide, as they significantly compromise animal welfare and increase production costs¹. The relationship between the parasite and livestock is influenced by several factors related to the host, namely the genetic characteristics of resistance and resilience of the animals, the nutritional status of the animals, the level of parasitism, and the pathogenicity of the parasite populations involved².

For years, parasite control has relied exclusiv....

Protocol

The CEMIT animal ethics committee approves all experiments involving animal manipulation. The egg-donor animals belong to farms that collaborate with CEMIT by providing fecal samples for preventive diagnostic purposes. These animals are sheep or goats (no specific breed), of either sex, over one year of age, and must not have been dewormed within 15 days prior to sample collection. All participating farms are informed about the use of the samples for research purposes and provide their consent to be part of the study. For egg collection, nearby establishments are first notified, and upon receiving permission, field visits are organized to collect fresh fecal samples, ....

Results

Total phenolic content (TPC), total flavonoid content (TFC), and total tannin content (TTC)
Results of TPC, TTC, TFC, and ABTS on Tithonia diversifolia are shown in Table 1. Values are expressed as mg of gallic acid equivalents per gram of dry matter (mg GAE/g DM) for TPC, mg of quercetin equivalents per gram of dry matter (mg QE/g DM) for TFC, and mg of catechin equivalents per gram of dry matter (mg CE/g DM) for TTC. Antioxidant capacity is expressed as the half-maximal in.......

Discussion

This article outlines several essential steps for advancing research on medicinal plants. In vitro tests, such as the egg hatch assay (EHA), are particularly attractive because they can be conducted under controlled laboratory conditions. The EHA is designed to evaluate the activity of plant extracts at various concentrations on nematode eggs, comparing the outcomes against both positive and negative controls. The results obtained offer valuable insights into the potential of plant-derived compounds for developi.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
15 mL tubes			On site
48-hole plate			On site
96 well plates	VWR	734-0126
ABTS	Enzymatic	A1088.0005	On site
Academic optical microscope and inverted microscope			On site
Acetone	Enzymatic	A/0600/17
Aluminum chloride	Sigma Aldrich	6911
DMACA reagent	Sigma Aldrich	49825
Ethanol 96%	Enzymatic	VR11202
Feces from animals			Collected in farms near to university
Folin Ciocalteau	Sigma Aldrich	F9252
Gallic acid	Sigma Aldrich	91215
HCl	Sigma Aldrich	258148
McMaster camera			On site
Methanol	Enzymatic	M/4000/17
PBS			On site
Plant extracts			On site
Potassium persulfate	Sigma Aldrich	216224
Quercetin	Sigma Aldrich	Q4951
Salt			On site
Sodium carbonate	Sigma Aldrich	1063920500
Thiabendazole			On site
Trolox	Sigma Aldrich	648471	On site
Water			On site
Whatmann no. 5	VWR	WHAT97039654