Isorhamnetin: A nematocidal flavonoid from prosopis laevigata leaves against haemonchus contortus eggs and larvae

Haemonchus contortus affect small ruminants all over the world. Anthelmintics cause resistance, contamination, and a risk of public health. Prosopis laevigata is a plant used as a home remedy against many diseases in Mexico. This study arose from a preliminary study where a P. laevigata hydroalcoholic extract (Pl-hae) showed anthelmintic activity (aa) against H. contortus. Searching for bioactive compounds (bac) with high aa, the Pl-hae was fractioned obtaining an aqueous (Aq-F) and an ethyl acetate fraction (EtAc-F), and a flavonoid with aa identified as isorhamnetin was obtained from EtAc-F. Both fractions were in vitro assessed by the egg hatch test (eht) and larval mortality (lm) assays. The bac obtained from EtAc-F were characterised by NMR analysis. The highest aa were recorded with EtAc-F, resulting in 100% eht and 80.45% lm at 0.75 and 30 mg/mL, respectively. Alterations in eggs and larvae attributed to isorhamnetin were recorded by environmental scanning electron microscopy, confocal laser scanning and by high-resolution digital-coupled camera. This flavonoid caused 100% eht at 0.07 mg/mL after 48 h and 100% lm at 7.5 mg/mL after 72 h exposure. Isorhamnetin has promising potential as an anthelmintic against sheep haemonchosis.