ABSTRACT
The number of Neospora in tissues of chronically infected animals is low, and tissue cysts are more likely to be found than tachyzoites. Numerous mammalian cell lines have been used to grow N. caninum in cell cultures and the procedures used to isolate viruses in diagnostic laboratories are suitable to grow N. caninum. In vitro culture of N. caninum tachyzoites has been established in a variety of primary cells, and also established cell lines. N. caninum infection was also studied in human brain microvascular endothelial cells. The number of Neospora oocysts in feces is usually low; therefore, concentration methods are often necessary to detect oocysts in feces. During collection and manipulation, DNA-free equipment should be used and cross-contamination and carry-over needs to be avoided and controlled using negative processing controls. Prolonged fixation with 4% or 10% formaldehyde buffers prior to paraffin embedding may impair a subsequent polymerase chain reaction analysis.
