Characterization of an alpha tubulin gene sequence from Neospora caninum and Hammondia heydorni, and their comparison to homologous genes from Apicomplexa

The gene coding for α tubulin has been isolated by the polymerase chain reaction and sequenced from 2 isolates of Neospora caninum (Nc-Liverpool and Nc-SweB1). The data show that the gene, as in Toxoplasma gondii, is single copy and contains 3 exons and 2 introns and is identical in sequence in the 2 isolates studied. Comparison of the predicted protein sequence shows it to be identical to the α tubulin protein encoded by the T. gondii gene. The majority of the nucleotide substitutions that have occurred during the evolution of the T. gondii and N. caninum genes from their common ancestor have occurred in the third codon position. A partial coding sequence for α tubulin was also obtained from Hammondia heydorni and compared to other α tubulin sequences from Apicomplexa. The results show the sequences of the T. gondii, N. caninum and H. heydorni α tubulin genes to be similar but not identical in sequence, thereby providing new evidence that N. caninum and H. heydorni are genetically distinct species

Genetic diversity amongst isolates of Neospora caninum, and the development of a multiplex assay for the detection of distinct strains

Infection with Neospora caninum is regarded as a significant cause of abortion in cattle. Despite the economic impact of this infection, relatively little is known about the biology of this parasite. In this study, mini and microsatellite DNAs were detected in the genome of N. caninum and eight loci were identified that each contained repetitive DNA which was polymorphic among different isolates of this parasite. A multiplex PCR assay was developed for the detection of genetic variation within N. caninum based on length polymorphism associated with three different repetitive markers. The utility of the multiplex PCR was demonstrated in that it was able to distinguish amongst strains of N. caninum used as either vaccine or challenge strains in animal vaccination experiments and that it could genotype N. caninum associated with naturally acquired infections of animals. The multiplex PCR is simple, rapid, informative and sensitive and should provide a valuable tool for further studies on the epidemiology of N. caninum in different host species