Proline modulates the effect of bisphosphonate on calcium levels and adenosine triphosphate production in cell lines derived from bovine Echinococcus granulosus protoscoleces A.G. Fuchs*, C.I. Echeverrı´a, F.G. Pe ´ rez Rojo, E.A. Prieto Gonza ´lez and E.J.A. Rolda ´n Centro de Altos Estudios en Ciencias Humanas y de la Salud (CAECIHS), Universidad Abierta Interamericana (UAI), Avenida Montes de Oca 745, (C127OAAH), Buenos Aires, Argentina (Received 20 September 2012; Accepted 28 April 2013; First Published Online 7 June 2013) Abstract Bisphosphonates have been proposed as pharmacological agents against parasite and cancer cell growth. The effect of these compounds on helminthic cell viability and acellular compartment morphology, however, has not yet been studied. The effects of different types of bisphosphonates, namely etidronate (EHDP), pamidronate (APD), alendronate (ABP), ibandronate (IB) and olpadronate (OPD), and their interaction with amiloride, 1,25-dihydroxychole- calciferol (D 3 ) and proline were evaluated on a cell line derived from bovine Echinococcus granulousus protoscoleces (EGPE) that forms cystic colonies in agarose. The EGPE cell line allowed testing the effect of bisphosphonates alone and in association with other compounds that could modulate calcium apposition/deposition, and were useful in measuring the impact of these compounds on cell growth, cystic colony formation and calcium storage. Decreased cell growth and cystic colony formation were found with EHDP, IB and OPD, and increased calcium storage with EHDP only. Calcium storage in EGPE cells appeared to be sensitive to the effect of amiloride, D 3 and proline. Proline decreased calcium storage and increased colony formation. Changes in calcium storage may be associated with degenerative changes of the cysts, as shown in the in vitro colony model and linked to an adenosine triphosphate (ATP) decrease. In conclusion, bisphosphonates could be suitable tempering drugs to treat cestode infections. Introduction Echinococcus granulosus and its genotypic variants are the aetiologic agents of cystic hydatid or echinococcal disease, which occurs widely in the American continent from Alaska and northern Canada to Tierra del Fuego (Moro & Schantz, 2006) and is caused by the metacestode stage of the tapeworm. Metacestodes are fluid-filled cysts with cellular and acellular compartments. The outer, acellular, layer of the metacestode is formed by a laminated membrane which becomes surrounded by a very prominent, host-derived fibrous capsule or adventi- tial layer (Hemphill et al., 2007). Spontaneous cure of hydatidosis occurs sometimes; it starts with a process of calcification of the laminated membrane. Bisphosphonates *Fax: (54) 11 4301 5240 extension 2 E-mail: Alicia.Fuchs@uai.edu.ar Journal of Helminthology (2014) 88, 459–467 doi:10.1017/S0022149X13000436 q Cambridge University Press 2013