âTachyzoites were detected in the flasks 1 week post infection (Fig. 2 A). âHP lessions in mice: mononuclear and neutrophil infiltrate in liver, heart and lungs.
NEOSPORA CANINUM ISOLATION FROM BEEF CATTLE: INMUNODIAGNOSTIC AND PCR STUDIES CAMPERO, L. M. 1, 2; DELLARUPE, A. 1, 2 ; GUAL, I. 3; MORÉ, G. A. 1, 2; RAMBEAUD, M. 1, 2; MORRELL, E. 3; 1 2 MOORE, D. P. ; VENTURINI, M.C . 1
Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina 2 Laboratorio de Inmunoparasitología, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, Argentina 3 Instituto Nacional de Tecnología Agropecuaria (INTA), Balcarce, CC 276, 7620 Balcarce, Argentina
Introduction
Results Dam, placenta and fetus
Bovine neosporosis is a parasitic disease caused by the protozoan Neospora caninum, manifested mainly by abortion between 4- 8 months of gestation, that causes significant economic losses in beef and dairy cattle.
Histopathological lesions compatible with N. caninum (miocarditis, meningoencephalitis and hemorrhage, lymphoplasmahistiocytic placentitis).
Objectives The aims of this work were to obtain isolates of N. caninum from a beef herd in
Clusters of tachyzoites were detected in the placenta by IHC PCR results were positive for the placenta and the CNS of the fetus (Fig. 1)
MW
Negative control
Positive control
Placenta
Dams CNS
Materials and methods
Fetus CNS
Argentina and to confirm diagnosis by immunodiagnostic and PCR analysis.
Fig. 1. PCR results on dams-fetus tissues
IFAT results: Serum-dam/ fetal liquid
1:3200
Mice 25 d.p.i. serum from 3 mice A 10 years old Angus cow with 6 months of gestation
(inoculated with CNS from fetus)
+ IFAT
Brains from seropositive mice were used for successive in vivo
Necropsied
passages and peritoneal washings were used to infect cell culture Tachyzoites were detected in the flasks 1 week post infection (Fig. 2 A) HP lessions in mice: mononuclear and neutrophil infiltrate in liver, heart and lungs
Central nervous system (CNS) from mother
Central nervous system (CNS) from fetus
Placenta
Digestion: 0,5 % trypsin Tissues were inoculated in gamma knockout mice
IHC positive staining in mice: lungs and heart (Fig. 2 B) Infected mice tissues (CNS, heart, lungs, liver and peritoneal fluid) and cell culture derived tachyzoites were positive by PCR A
B
Fig. 2. A: Tachyzoites in mouse peritoneal washing; B: Positive IHC in infected mouse heart
Discussion Multilocus microsatellite analysis is currently underway to genetically characterize this new bovine isolate. Further bioassays will be performed to characterize this new isolate. Histopathology (HP) Immunohistochemistry (IHC) Polymerase chain reaction (PCR) Indirect fluorescent antibody test (IFAT)
References (1) Dubey JP, Schares G, Ortega-Mora LM. Epidemiology and control of neosporosis and Neospora caninum. Clin Microbiol Rev2007 Apr 20(2):323-67 (2) Schares, G., Peters, M., Wurm, R., Bärwald, A. and Conraths, F.J. (1998) The efficiency of vertical transmission of Neospora caninum in dairy cattle analysed by serological techniques. Veterinary Parasitology 80, 87–98
