Genetic Diversity and Geographic Population Structure of Bovine Neospora caninum Determined by Microsatellite Genotyping Analysis

Abstract

The cyst-forming protozoan parasite Neospora caninum is one of the main causes of bovine abortion worldwide and is of great economic importance in the cattle industry. Recent studies have revealed extensive genetic variation among N . caninum isolates based on microsatellite sequences (MSs). MSs may be suitable molecular markers for inferring the diversity of parasite populations, molecular epidemiology and the basis for phenotypic variations in N . caninum , which have been poorly defined. In this study, we evaluated nine MS markers using a panel of 11 N . caninum -derived reference isolates from around the world and 96 N . caninum bovine clinical samples and one ovine clinical sample collected from four countries on two continents, including Spain, Argentina, Germany and Scotland, over a 10-year period. These markers were used as molecular tools to investigate the genetic diversity, geographic distribution and population structure of N . caninum . Multilocus microsatellite genotyping based on 7 loci demonstrated high levels of genetic diversity in the samples from all of the different countries, with 96 microsatellite multilocus genotypes (MLGs) identified from 108 N . caninum samples. Geographic sub-structuring was present in the country populations according to pairwise F ST. Principal component analysis (PCA) and Neighbor Joining tree topologies also suggested MLG segregation partially associated with geographical origin. An analysis of the MLG relationships, using eBURST, confirmed that the close genetic relationship observed between the Spanish and Argentinean populations may be the result of parasite migration (i.e., the introduction of novel MLGs from Spain to South America) due to cattle movement. The eBURST relationships also revealed genetically different clusters associated with the abortion. The presence of linkage disequilibrium, the co-existence of specific MLGs to individual farms and eBURST MLG relationships suggest a predominant clonal propagation for Spanish N . caninum MLGs in cattle.

All about neosporosis in Brazil

“…Additionally, the Brazilian NC-Bahia strain isolated from the dog was included among the N. caninum strains characterized by researchers outside Brazil (AL-QASSAB et al, 2009, 2010REGIDOR-CERRILLO et al, 2013).…”

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“…Neospora caninum genotyping DNA samples obtained from N. caninum PCR-positive brains were used for genotyping by multilocus microsatellite analysis. Specifically, MS4, MS5, MS6A, MS6B, MS7, MS8, MS10, MS12 and MS21 markers were amplified using specific primers and nested-PCR conditions as described by Regidor-Cerrillo et al [20]. The sizes of the PCR products for all microsatellites were determined in a 48-capillary 3730 DNA Analyser (Applied Biosystems, Foster City, CA, USA) with GeneScan-500 (LIZ) size standards (Applied Biosystems) (at the Unidad Genómica del Parque Científico de Madrid), and the results were analyzed with GeneMapper1 v3.5 software.…”

Reevaluation of theToxoplasma gondiiandNeospora caninumgenomes reveals misassembly, karyotype differences and chromosomal rearrangements

“…This is consistent with our results whereby our whole genome assembly of NcUru1 is practically indistinguishable from that of NcLiv, despite deriving from completely different geographical locations (Europe vs. South America). Nonetheless, it is well established that great genetic variability exists amongst N. caninum strains in the form of SNPs at particular loci and that these genetic differences underlie phenotypic variability 12,51,52 . In this context, it is noteworthy that linkage assays, whereby these minimal differences can be correlated to virulence phenotypes, rely on correct genome assemblies.…”