Genetic Diversity of Neospora Caninum in Palestinian Cattle and Sheep: Insights From Nc‐5 Gene Detection and MS10 Microsatellite Analysis

INTRODUCTION: Neosporosis is an infectious disease responsible for abortions in dairy cattle worldwide. This study aimed to detect Neospora caninum (N. caninum) in cattle and sheep in Palestine and to characterise the identified isolates genetically. METHODS: A total of 124 brain tissue specimens were obtained from slaughtered animals at Jericho abattoir, comprising 106 cattle (85%) and 18 sheep (15%). Polymerase chain reaction (PCR) targeting the Nc-5 gene and DNA Sanger sequencing were performed sequentially for the detection of N. caninum. The genetic variation and population structure were investigated using the MS10 microsatellite marker. RESULTS: Neospora caninum DNA was detected in 24.2% (30/124) of samples, with cattle showing a higher infection rate (25.5%) compared to sheep (16.7%). DNA sequencing of 17 randomly selected positive samples confirmed the infection. Phylogenetic and genetic clustering analyses, incorporating 309 global N. caninum samples, including 17 from Palestine, revealed minimal genetic diversity and indicated the presence of two main clusters that were not associated with either geographical origin or host species. The Palestinian isolates are highly monomorphic, forming a single genetic cluster with three MS10 alleles, of which the ACT allele predominated (76%). CONCLUSION: This study successfully identified and characterised N. caninum in cattle and sheep from Palestine compared to the worldwide available MS10 repeats. These findings contribute to the broader understanding of N. caninum distribution and genetic diversity. Based on our study findings, we recommend implementing routine surveillance programmes for N. caninum in livestock to enable early detection and reduce economic losses.