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Despite the importance of\n                    <jats:italic>Babesia<\/jats:italic>\n                    in livestock industry and emerging cases in humans, their basic biology is hampered by lack of suitable biological tools. In this study, we aimed to develop a synchronization method for\n                    <jats:italic>Babesia bovis<\/jats:italic>\n                    which causes the most pathogenic form of bovine babesiosis. Initially, we used compound 2 (C2), a specific inhibitor of cyclic GMP-dependent protein kinase (PKG), and a derivative of C2, ML10. While both inhibitors were able to prevent\n                    <jats:italic>B. bovis<\/jats:italic>\n                    egress from RBC and increased percentage of binary forms, removal of inhibitors from culture did not result in a synchronized egress of parasites. Because using PKG inhibitors alone was not efficient to induce a synchronized culture, we isolated viable and invasive\n                    <jats:italic>B. bovis<\/jats:italic>\n                    merozoites and showed dynamics of merozoite invasion and development in RBCs. Using isolated merozoites we showed that BbVEAP, VESA1-export associated protein, is essential for parasite development in the RBC while has no significant role in invasion. Given the importance of invasion for the establishment of infection, this study paves the way for finding novel antigens to be used in control strategies against bovine babesiosis.\n                  <\/jats:p>","DOI":"10.1038\/s41598-021-96365-w","type":"journal-article","created":{"date-parts":[[2021,8,20]],"date-time":"2021-08-20T06:03:20Z","timestamp":1629439400000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Isolation of viable Babesia bovis merozoites to study parasite invasion"],"prefix":"10.1038","volume":"11","author":[{"given":"Hassan","family":"Hakimi","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masahito","family":"Asada","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takahiro","family":"Ishizaki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shinichiro","family":"Kawazu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2021,8,20]]},"reference":[{"key":"96365_CR1","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/S0065-308X(08)60285-7","volume":"23","author":"H Mehlhorn","year":"1984","unstructured":"Mehlhorn, H. & Shein, E. The piroplasms: Life cycle and sexual stages. Adv. Parasitol. 23, 37\u2013103 (1984).","journal-title":"Adv. Parasitol."},{"key":"96365_CR2","doi-asserted-by":"publisher","first-page":"248","DOI":"10.3389\/fcimb.2018.00248","volume":"8","author":"M Jalovecka","year":"2018","unstructured":"Jalovecka, M., Hajdusek, O., Sojka, D., Kopacek, P. & Malandrin, L. The complexity of piroplasms life cycles. Front. Cell. Infect. Microbiol. 8, 248 (2018).","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"96365_CR3","first-page":"1","volume":"00","author":"B Elsworth","year":"2020","unstructured":"Elsworth, B. & Duraisingh, M. T. A framework for signaling throughout the life cycle of Babesia species. Mol. Microbiol. 00, 1\u20139 (2020).","journal-title":"Mol. Microbiol."},{"issue":"4436","key":"96365_CR4","doi-asserted-by":"publisher","first-page":"1218","DOI":"10.1126\/science.7355284","volume":"207","author":"MG Levy","year":"1980","unstructured":"Levy, M. G. & Ristic, M. Babesia bovis: Continuous cultivation in a microaerophilous stationary phase culture. Science 207(4436), 1218\u20131220 (1980).","journal-title":"Science"},{"key":"96365_CR5","doi-asserted-by":"publisher","first-page":"269","DOI":"10.3389\/fcimb.2020.00269","volume":"10","author":"MJ Gubbels","year":"2020","unstructured":"Gubbels, M. J. et al. Fussing about fission: Defining variety among mainstream and exotic apicomplexan cell division modes. Front. Cell. Infect. Microbiol. 10, 269 (2020).","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"96365_CR6","doi-asserted-by":"publisher","first-page":"859","DOI":"10.1111\/cmi.12555","volume":"18","author":"JR Cursino-Santos","year":"2016","unstructured":"Cursino-Santos, J. R., Singh, M., Pham, P., Rodriguez, M. & Lobo, C. A. Babesia divergens builds a complex population structure composed of specific ratios of infected cells to ensure a prompt response to changing environmental conditions. Cell Microbiol. 18, 859\u2013874 (2016).","journal-title":"Cell Microbiol."},{"key":"96365_CR7","doi-asserted-by":"publisher","first-page":"851","DOI":"10.1111\/j.1462-5822.2006.00834.x","volume":"9","author":"A Alkhalil","year":"2007","unstructured":"Alkhalil, A., Hill, D. A. & Desai, S. A. Babesia and plasmodia increase host erythrocyte permeability through distinct mechanisms. Cell Microbiol. 9, 851\u2013860 (2007).","journal-title":"Cell Microbiol."},{"issue":"10","key":"96365_CR8","doi-asserted-by":"publisher","first-page":"e1008917","DOI":"10.1371\/journal.ppat.1008917","volume":"16","author":"H Hakimi","year":"2020","unstructured":"Hakimi, H. et al. Novel\u00a0Babesia bovis\u00a0exported proteins that modify properties of infected red blood cells. 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