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Neosporosis is considered as one of the main causes of abortion and severe economic losses in dairy industry. The  Canis  genus serving as one of the confirmed definitive hosts of the apicomplexan parasite  Neospora caninum  ( N. caninum ) plays a critical role in its life cycle. However, the effects of  N. caninum  on its definitive hosts of neutrophils extracellular traps (NETs) formation remain unclear. In the present study,  N. caninum  tachyzoite-induced canine NETs formation was observed by scanning electron microscopy (SEM). Visualization of DNA decorated with H3, neutrophil elastase (NE), and myeloperoxidase (MPO) within  N. caninum  tachyzoite-induced NETs were examined using fluorescence confocal microscopy analyses. Furthermore, the formation of canine NETs was quantified using Sytox Green staining, and the LDH levels in supernatants were examined by an LDH Cytotoxicity Assay ®  kit. The results clearly showed that NETs-like structures were induced by  N. caninum  tachyzoites, and the major components within these structures induced by  N. caninum  tachyzoite were further confirmed by fluorescence confocal microscopy visualization. These results suggest that  N. caninum  tachyzoites strongly induced NETs formation in canine polymorphonuclear neutrophils (PMN). In functional inhibition assays, the blockings of NADPH oxidase, NE, MPO, SOCE, ERK 1/2, and p38 MAPK signaling pathways significantly inhibited  N. caninum  tachyzoite-induced NETs formation. To our knowledge, this study is the first to report the formation of NETs in canine PMN against  N. caninum  infection.

Canine Neutrophil Extracellular Traps Release Induced by the Apicomplexan Parasite Neospora caninum In Vitro