In vitro anthelmintic effect of Vicia pannonica var. purpurascens on trichostrongylosis in sheep Esma Kozan a,⇑ , Serap Arabaci Anul b , I. Irem Tatli b,⇑ a Department of Parasitology, Faculty of Veterinary Medicine, Afyon Kocatepe University, 03200 Afyonkarahisar, Turkey b Department of Pharmaceutical Botany, Faculty of Pharmacy, Hacettepe University, Sihhiye, 06100 Ankara, Turkey highlights  Vicia species are used in Turkish traditional medicine.  We examined the in vitro anthelmintic effects of Vicia pannonica extracts.  This plant possesses a significant in vitro anthelmintic activity. graphical abstract The acetone extract of Vicia pannonica var. purpurascens and its flavonoid glucosides possesses a signifi- cant in vitro anthelmintic activity which support its traditional utilization. article info Article history: Received 26 September 2012 Received in revised form 12 February 2013 Accepted 18 March 2013 Available online 4 April 2013 Keywords: Fabaceae Flavonoid In vitro test Trichostrongylosis Vicia abstract Vicia species are used for the treatment of malaria, diarrhea, hemorrhoids, kidney problems and infertility in Turkish traditional medicine. The present study was carried out to evaluate the in vitro anthelmintic effects of Vicia pannonica Crantz. var. purpurascens (DC). Ser. extracts. Larval motility test was used to determine anthelmintic activity of this plant. Motility of the larvae is measured by observation. The methanol, n-hexane, chloroform, acetone, and aqueous extracts of the aerial parts of the plant including the leaves and flowers were applied to developing trichostrongylus larvae at 1, 0.8, 0.6, 0.4, 0.2 and 0.1 mg/ml doses. Thiabendazole and distilled water with 5% DMSO was used as positive and negative control. All of the extracts were 100% effective. Two flavone and flavonol glycosides; luteolin-7-b-O- glucopyranoside (1) and quercetin-3-O-b-glucopyranoside (2) were isolated from the acetone extract and their structures were elucidated by spectral techniques. The solutions prepared from two flavonoid fractions at several doses were performed in vitro to larvae in the same way. Both of them were 100% effective at 1 and 0.8 mg/ml doses. Results of the present study support the utilization of these plant spe- cies employed in Turkish folk medicine. Ó 2013 Elsevier Inc. All rights reserved. 1. Introduction Gastrointestinal parasites are recognized as major problems to livestock production in tropical and developing countries (Ade- jinmi and Harrison, 1997; Hounzangbe-Adote et al., 2005). The economic impacts of these parasites, as production diseases in ruminants lie not only in direct losses such as mortality associated with the clinical forms of diseases but also indirect insidious losses as a result of weaknesses (Gibbs, 1986). In generally modern synthetic anthelmintics are used in control of these parasites (Kaplan, 2004; Coles et al., 2006). However helminth control programs, base on improvement of farm manage- ment and regular anthelmintic treatment, are often impracticable in developing countries because of relatively high price of modern anthelmintics for smallholder (Satrija et al., 2001). In addition, the development of anthelmintic resistance in nematodes has been re- ported throughout the world (Waller, 1994; Jackson and Coop, 2000). Furthermore some drugs may cause food residues and envi- ronmental pollution (Hammond et al., 1997). For these reasons, re- cently, the new alternative methods or nonchemical agents have been needed. A variety of in vitro assays have been developed for anthelmintic properties, in which the parasite stages are directly incubated in the chemical compound (Taylor et al., 2002). Larval motility assays measure the ability of anthelmintic to paralyse the infective third larval stage. Therefore, the overall principle of these assays relies on the assessment of larvae motility. The larval 0014-4894/$ - see front matter Ó 2013 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.exppara.2013.03.018 ⇑ Corresponding authors. Fax: +90 312 3114777. E-mail addresses: esmakozan@aku.edu.tr (E. Kozan), itatli@hacettepe.edu.tr (I.I. Tatli). Experimental Parasitology 134 (2013) 299–303 Contents lists available at SciVerse ScienceDirect Experimental Parasitology journal homepage: www.elsevier.com/locate/yexpr