Abstract
In the present work, the anthelmintic activity (AA) of ethanolic extracts obtained from Gliricidia sepium, Leucaena leucocephala, and Pithecellobium dulce was evaluated using the third-stage-larval (L3) exsheathment inhibition test (LEIT) and egg hatch test (EHT) on Haemonchus contortus. Extracts were tested at concentrations of 0.3, 0.6, 1.2, 2.5, 5.0, 10, 20, and 40 mg/mL. The larval exsheathment inhibition (LEI) results showed that G. sepium achieved the highest average inhibition of 91.2%, compared with 44.6% for P. dulce and 41.0% for L. leucocephala at a concentration of 40 mg/mL; the corresponding IC50 values were 22.4, 41.7, and 43.3 mg/mL, respectively. The rates of egg hatching inhibition (EHI) at a concentration of 5 mg/mL were 99.5% for G. sepium, 64.2% for P. dulce, and 54% for L. leucocephala; the corresponding IC50 values were 1.9 mg/mL for G. sepium, 3.9 mg/mL for P. dulce, and 4.3 mg/mL for L. leucocephala. The species extracts studied here were also analyzed by ultra-high performance liquid chromatography and Orbitrap high resolution mass spectrometry (UHPLC-Q/Orbitrap/MS/MS), resulting in the compounds’ identification associated with AA. Glycosylated flavonoids and methoxyphenols were observed in all three species: fatty acids in G. sepium and P. dulce; phenylpropanoids, anthraquinone glycosides, amino acids and glycosylated phenolic acids in G. sepium; and flavonoids in L. leucocephala. Comparatively, G. sepium presented a greater diversity of compounds potentially active against the control of gastrointestinal nematodes, which was associated with the results obtained in the applied tests.
Highlights
Gastrointestinal parasitism is among the factors with the greatest adverse economic impact on the world’s ovine industry [1,2]
Compounds with an anthelmintic effect were identified as glycosylated flavonoids and methoxyphenols in all three species: fatty acids in G. sepium and P. dulce; phenylpropanoids other than flavonoids, anthraquinonic glycosides, amino acids, and glycosylated phenolic acids in G. sepium; and flavonoids in L. leucocephala
In order to evaluate the in vitro anthelmintic activity (AA) of the extracts of leaves of G. sepium, L. leucocephala, and P. dulce, LEIT and IHT were applied, and the compounds with potential AA were identified by analysis of the extracts using UHPLC-Q/Orbitrap/MS/MS
Summary
Gastrointestinal parasitism is among the factors with the greatest adverse economic impact on the world’s ovine industry [1,2]. Parasites of the Trichostrongylidae, Strongylidae, Trichuridae, and Ancylostomatidae families were found at prevalence rates of 92.5%, 48.3%, 12.0%, and 11.0% in sheep in Tolima, Colombia [7]. Attempts at controlling gastrointestinal parasitism by using chemical substances indiscriminately without epidemiological knowledge [9] has given rise to anthelmintic resistance and chemical traces being present in both food and the environment [10]. This anthelmintic resistance has been demonstrated in different parts of the world [11,12,13,14,15,16]. In Colombia, multiple resistance in gastrointestinal nematodes (H. contortus, Trichostrongylus spp., and Cooperia spp.) has been found, in the regions of Cundinamarca and Valle del Cauca [17]
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