Short communication Confirmed clinical Neospora caninum infection in a boxer puppy from Argentina W. Basso a , M.C. Venturini a, * , D. Bacigalupe a , M. Kienast b , J.M. Unzaga a , A. Larsen a , M. Machuca c , L. Venturini a a Laboratorio de Inmunoparasitologı ´a, Ca ´tedra de Parasitologı ´a y Enfermedades Parasitarias, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, 60 y 118, 1900 La Plata, Argentina b Ca ´tedra de Gene ´tica y Biometrı ´a, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, 60 y 118, 1900 La Plata, Argentina c Ca ´tedra de Patologı ´a Especial, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, 60 y 118, 1900 La Plata, Argentina Received 22 March 2005; received in revised form 29 March 2005; accepted 6 May 2005 Abstract Generalized neosporosis was diagnosed in a 2-month-old boxer puppy. The dog had a history of progressive paralysis and muscle atrophy, followed by cervical weakness, stiff jaws and dysphagia. The dog had a 1:12,800 antibody titer for Neospora caninum and was negative for antibodies to Toxoplasma gondii by the indirect fluorescent antibody test (IFAT). After euthanasia a complete necropsy was carried out. The puppy had a megaesophagus. Microscopically, tachyzoites and tissue cysts were observed in histologic brain sections. Severe myositis was observed in esophagus and striated muscle sections and several groups of tachyzoites were associated with these lesions. Immunohistochemically, parasites in the brain and striated muscle reacted to anti-N. caninum antiserum. Western blot analysis allowed the identification of three major and four minor antigens of N. caninum tachyzoites corresponding to 30, 37, 45-kDa and 28, 29, 43, 47 and 67-kDa bands, respectively. Cerebral homogenate of the dog was inoculated into four Mongolian gerbils (Meriones unguiculatus). Forty-nine days after inoculation, all the gerbils had positive IFAT titers to N. caninum (1:200, 1:400, 1:100 and 1:400). Genomic DNA was isolated from the brain, lung and striated muscle from the puppy and from the brain of one of the inoculated gerbils. The N. caninum specific primer pair Np 6/21 produced 328 bp amplicons on electrophoretic gels. This is the first confirmed clinical case of generalized canine neosporosis in Argentina. # 2005 Elsevier B.V. All rights reserved. Keywords: Neospora caninum; Dog; Immunohistochemistry; Western blot; PCR; Gerbils; Argentina 1. Introduction Neospora caninum is a protozoan parasite that causes abortion in cows and neurological disorders www.elsevier.com/locate/vetpar Veterinary Parasitology 131 (2005) 299–303 * Corresponding author. Tel.: +54 221 4249621; fax: +54 221 4257980. E-mail address: cventuri@fcv.unlp.edu.ar (M.C. Venturini). 0304-4017/$ – see front matter # 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.vetpar.2005.05.003