Isolation of Neospora caninum from kidney and brain of a bovine foetus and molecular characterization in Brazil Rosangela Locatelli Dittrich a, * , Javier Regidor-Cerrillo b , Luis Miguel Ortega-Mora b , Marília de Oliveira Koch a , Ana Paula B. Busch a , Kamila Alcal  a Gonçalves a , Amilcar A. Cruz c a Programa de P os Graduaç~ ao em Ci^ encias Veterin arias, Setor de ci^ encias Agr arias, Universidade Federal do Paran a, Rua dos Funcion arios, 1540, CEP 80035- 050, Curitiba, Paran a, Brazil b Animal Health Department, SALUVET, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain c Universidade de Camagüey, Camagüey, Cuba highlights  N. caninum can be isolated from kidney of a foetus.  The new isolate, BNC-PR4 is a bovine isolate with low virulence.  Multilocus-microsatellite genotyping revealed a unique genetic profile that differed from previously reported isolates.  We report the first bovine isolate of N. caninum from kidney. article info Article history: Received 7 August 2017 Received in revised form 11 October 2017 Accepted 2 January 2018 Available online 5 January 2018 Keywords: Neospora caninum Isolate Kidney Microsatellite typing abstract Bovine neosporosis has become a disease of international concern as it is among the main causes of abortion in cattle. Viable N. caninum has been isolated from brains of fetuses and neonatal calves, and there is no report of isolation of tachyzoites from kidney. Also, detailed information about the genetic diversity of N. caninum is scarce. N. caninum tachyzoites were isolated from the kidney and the brain of an aborted 4-month-old bovine foetus. The parasite was confirmed to be N. caninum by PCR. The tachyzoites of the new isolate, named BNC-PR4, were propagated in Vero cell cultures. Pathogenicity of the parasite was examined in BALB/c mice. Mice inoculated intraperitoneally with BNC-PR4 failed to yield clinical signs of disease and did not induce severe brain lesions, suggesting a bovine isolate with low virulence. The N. caninum-positive DNA sample was further analyzed by multilocus microsatellite (MS) genotyping for MS4, MS5, MS6A, MS6B, MS7, MS8, MS10, MS12, and MS21. Multilocus- microsatellite genotyping revealed a unique genetic profile that differed from previously reported isolates. Published by Elsevier Inc. 1. Introduction Neospora caninum is a protozoan parasite of the phylum Api- complexa, and has been recognized as a major cause of abortion and reproductive failure in cattle, in many countries (Dubey and Schares, 2011; Campero et al., 2015). Congenital infection has been shown to be important in the spread of N. caninum and it appears to be the major mode of transmission in cattle (Hall et al., 2005). The efficiency of vertical transmission is important for the persistence of N. caninum within herds and can result in abortion, weak and underweight calves, or as in most cases, asymptomatic congenitally infected newborns (Dubey et al., 2007). Infection in a pregnant animal may result in abortion. World- wide, these abortions are a major root cause of economic loss to both the dairy and beef industries (Larson et al., 2004; Dubey and Schares, 2011; Reichel et al., 2013). In addition to abortions, N. caninum causes neurological syndrome in calves (Locatelli- Dittrich et al., 2003; Santos et al., 2006). In Brazil, N. caninum DNA was detected in the brain of adult cattle with neurological * Corresponding author. E-mail address: roslocdi@ufpr.br (R. Locatelli Dittrich). Contents lists available at ScienceDirect Experimental Parasitology journal homepage: www.elsevier.com/locate/yexpr https://doi.org/10.1016/j.exppara.2018.01.004 0014-4894/Published by Elsevier Inc. Experimental Parasitology 185 (2018) 10e16