Abstract
Neospora caninum is an apicomplexan protozoan and a major cause of abortion in cattle worldwide. In the Argentinian Humid Pampa, bovine neosporosis causes severe economic losses. Despite this, information on the genetic structure of N. caninum in this region is limited. Therefore, this study aimed to genetically characterize N. caninum isolates associated with bovine abortion in the Humid Pampa region. For this purpose, spontaneous bovine fetal tissues submitted for diagnosis to the Veterinary Diagnostic Service at INTA Balcarce during 2008–2019 were assessed by PCR, indirect fluorescent antibody test (IFAT), and histologic analysis. PCR-positive samples were tested by multilocus microsatellite genotyping (MLGs) using 9 microsatellite markers. Thirty-one different genotypes were identified from 32 samples with at least seven markers. Argentinian MLGs were grouped into two clonal clusters when analyzed using eBURST network and principal coordinate analysis. No segregation based on the year of collection, animal biotype, or geographic origin was observed. In addition, the presence of linkage disequilibrium supported the clonal propagation of Argentinian MLGs. One Argentinian subpopulation was associated with isolates from Spain, Uruguay, Brazil, and Mexico, and the other one was linked to isolates from Scotland, Spain, and Germany. These findings reveal the presence of two clonal subpopulations of N. caninum in the Humid Pampa.
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References
Anderson ML, Andrianarivo AG, Conrad PA (2000) Neosporosis in cattle. Anim Reprod Sci 60:417–431. https://doi.org/10.1016/S0378-4320(00)00117-2
Arelovich HM, Bravo RD, Martínez MF (2011) Development, characteristics, and trends for beef cattle production in Argentina. Anim Front 1:37–45. https://doi.org/10.2527/af.2011-0021
Basso W, Schares S, Minke L, Bärwald A, Maksimov A, Peters M, Schulze C, Müller M, Conraths FJ, Schares G (2010) Microsatellite typing and avidity analysis suggest a common source of infection in herds with epidemic Neospora caninum-associated bovine abortion. Vet Parasitol 173:24–31. https://doi.org/10.1016/j.vetpar.2010.06.009
Buxton D, Maley SW, Wright S, Thomson KM, Rae AG, Innes EA (1998) The pathogenesis of experimental neosporosis in pregnant sheep. J Comp Pathol 118:267–279. https://doi.org/10.1016/S0021-9975(07)80003-X
Cabrera A, Fresia P, Berná L, Silveira C, Macías-Rioseco M, Arevalo AP, Crispo M, Pritsch O, Riet-Correa F, Giannitti F, Francia ME, Robello C (2019) Isolation and molecular characterization of four novel Neospora caninum strains. Parasitol Res 118:3535–3542. https://doi.org/10.1007/s00436-019-06474-9
Campero CM, Anderson ML, Conosciuto G, Odriozola H, Bretschneider G, Poso MA (1998) Neospora caninum associated abortion in dairy herd in Argentina. Vet Rec 143:228–229. https://doi.org/10.1136/vr.143.8.228
Campero CM, Moore DP, Odeón AC, Cipolla AL, Odriozola E (2003) Aetiology of bovine abortion in Argentina. Vet Res Commun 27:359–369. https://doi.org/10.1023/A:1024754003432
Cappellini OR (2011) Dairy development in Argentina. FAO http://wwwfaoorg/3/al744e/al744e00pdf Accessed 22 May 2020
Davison HC, Otter A, Trees AJ (1999) Estimation of vertical and horizontal transmission parameters of Neospora caninum infections in dairy cattle. Int J Parasitol 29:1683–1689. https://doi.org/10.1016/S0020-7519(99)00129-0
Donahoe SL, Lindsay SA, Krockenberger M, Phalen D, Šlapeta J (2015) A review of neosporosis and pathologic findings of Neospora caninum infection in wildlife. Int J Parasitol Parasites Wildl 4:216–238. https://doi.org/10.1016/j.ijppaw.2015.04.002
Dubey JP, Schares G (2011) Neosporosis in animals-the last five years. Vet Parasitol 180:90–108. https://doi.org/10.1016/j.vetpar.2011.05.031
Dubey JP, Hattel AL, Lindsay DS et al (1988) Neonatal Neospora caninum infection in dogs: isolation of the causative agent and experimental transmission. J Am Vet Med A 193:1259–1263
Dubey JP, Buxton D, Wouda W (2006) Pathogenesis of bovine neosporosis. J Comp Pathol 134:267–289. https://doi.org/10.1016/j.jcpa.2005.11.004
Dubey JP, Schares G, Ortega-Mora LM (2007) Epidemiology and control of neosporosis and Neospora caninum. Clin Microbiol Rev 20:323–367. https://doi.org/10.1128/CMR.00031-06
FAOSTAT (2018) Statistical databases. http://www.fao.org/faostat/en/#data. Accessed 22 May 2020
Field D, Wills C (1996) Long, polymorphic microsatellites in simple organisms. Proc R Soc Lond B Biol Sci 263:209–215. https://doi.org/10.1098/rspb.1996.0033
Francisco AP, Bugalho M, Ramirez M, Carriço JA (2009) Global optimal eBURST analysis of multilocus typing data using a graphic matroid approach. BMC Bioinformatics 10:1–15. https://doi.org/10.1186/1471-2105-10-152
Guevara JC, Grunwaldt EG (2012) Status of beef cattle production in Argentina over the last decade and its prospects. In: Javed K (ed) Livestock production. IntechOpen, Khalid Javed, pp 117–134. https://doi.org/10.5772/50971
Haubold B, Hudson RR (2000) LIAN 3.0: detecting linkage disequilibrium in multilocus data. Linkage analysis. Bioinformatics. 16:847–848. https://doi.org/10.1093/bioinformatics/16.9.847
Hecker YP, Moore DP, Quattrocchi V, Regidor-Cerrillo J, Verna A, Leunda MR, Morrell E, Ortega-Mora LM, Zamorano P, Venturini MC, Campero CM (2013) Immune response and protection provided by live tachyzoites and native antigens from the NC-6 Argentina strain of Neospora caninum in pregnant heifers. Vet Parasitol 197(3–4):436–446. https://doi.org/10.1016/j.vetpar.2013.07.027
Khan A, Fujita AW, Randle N, Regidor-Cerrillo J, Shaik JS, Shen K, Oler AJ, Quinones M, Latham SM, Akanmori BD, Cleaveland S, Innes EA, Ryan U, Šlapeta J, Schares G, Ortega-Mora LM, Dubey JP, Wastling JM, Grigg ME (2019) Global selective sweep of a highly inbred genome of the cattle parasite Neospora caninum. Proc Natl Acad Sci U S A 116:22764–22773. https://doi.org/10.1073/pnas.1913531116
Kirkbride CA (1986) Examination of bovine and ovine fetuses. Vet Clin North Am Food Anim Pract 2:61–83. https://doi.org/10.1016/S0749-0720(15)31281-0
Lazzarini B, Baudracco J, Tuñon G et al (2019) Milk production from dairy cows in Argentina: current state and perspectives for the future. Appl Anim Sci 35:426–432. https://doi.org/10.15232/aas.2019-01842
Locatelli Dittrich R, Regidor-Cerrillo J, Ortega-Mora LM, Oliveira Koch M, Busch APB, Gonçalves KA, Cruz AA (2018) Isolation of Neospora caninum from kidney and brain of a bovine foetus and molecular characterization in Brazil. Exp Parasitol 185:10–16. https://doi.org/10.1016/j.exppara.2018.01.004
McAllister MM, Dubey J, Lindsay DS et al (1998) Dogs are definitive hosts of Neospora caninum. Int J Parasitol 28:1473–1479. https://doi.org/10.1016/S0020-7519(98)00138-6
Medina-Esparza L, Regidor-Cerrillo J, García-Ramos D, Álvarez-García G, Benavides J, Ortega-Mora LM, Cruz-Vázquez C (2016) Genetic characterization of Neospora caninum from aborted bovine foetuses in Aguascalientes, Mexico. Vet Parasitol 228:183–187. https://doi.org/10.1016/j.vetpar.2016.09.009
Moore D, Reichel M, Spath E, Campero C (2013) Neospora caninum causes severe economic losses in cattle in the humid pampa region of Argentina. Trop Anim Health Prod 45:1237–1241. https://doi.org/10.1007/s11250-013-0353-z
Morrell EL, Campero CM, Cantón GJ, Odeón AC, Moore DP, Odriozola E, Paolicchi F, Fiorentino MA (2019) Current trends in bovine abortion in Argentina. Pesq Vet Bras 39:12–19. https://doi.org/10.1590/1678-5150-pvb-5668
Nei M (1978) Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics 89:583–590
Oliveira S, Soares RM, Aizawa J et al (2017) Isolation and biological and molecular characterization of Neospora caninum (NC-SP1) from a naturally infected adult asymptomatic cattle (Bos taurus) in the state of São Paulo, Brazil. Parasitology 144:707–711. https://doi.org/10.1017/S0031182016002481
Payne S, Ellis J (1996) Detection of Neospora caninum DNA by the polymerase chain reaction. Int J Parasitol 26:347–351. https://doi.org/10.1016/0020-7519(96)00030-6
Peakall R, Smouse PE (2012) GenAlEx 6.5: genetic analysis in excel. Population genetic software for teaching and research–an update. Bioinformatics. 28:2537–2539. https://doi.org/10.1093/bioinformatics/bts460
Pedraza-Díaz S, Marugán-Hernández V, Collantes-Fernández E, Regidor-Cerrillo J, Rojo-Montejo S, Gómez-Bautista M, Ortega-Mora LM (2009) Microsatellite markers for the molecular characterization of Neospora caninum: application to clinical samples. Vet Parasitol 166:38–46. https://doi.org/10.1016/j.vetpar.2009.07.043
Poli MA, Antonini AG (1991) Genetic structure of milk proteins in Argentinian holstein and Argentinian creole cattle. Hereditas 115:177–182
Regidor-Cerrillo J, Pedraza-Díaz S, Gómez-Bautista M, Ortega-Mora LM (2006) Multilocus microsatellite analysis reveals extensive genetic diversity in Neospora caninum. J Parasitol 92:517–524. https://doi.org/10.1645/GE-713R.1
Regidor-Cerrillo J, Díez-Fuertes F, García-Culebras A, Moore DP, González-Warleta M, Cuevas C, Schares G, Katzer F, Pedraza-Díaz S, Mezo M, Ortega-Mora LM (2013) Genetic diversity and geographic population structure of bovine Neospora caninum determined by microsatellite genotyping analysis. PLoS One 8:1–12. https://doi.org/10.1371/journal.pone.0072678
Regidor-Cerrillo J, Horcajo P, Ceglie L, Schiavon E, Ortega-Mora LM, Natale A (2020) Genetic characterization of Neospora caninum from Northern Italian cattle reveals high diversity in European N. caninum populations. Parasitol Res 119:1353–1362. https://doi.org/10.1007/s00436-020-06642-2
Rojo-Montejo S, Collantes-Fernández E, Regidor-Cerrillo J, Álvarez-García G, Marugan-Hernández V, Pedraza-Díaz S, Blanco-Murcia J, Prenafeta A, Ortega-Mora LM (2009) Isolation and characterization of a bovine isolate of Neospora caninum with low virulence. Vet Parasitol 159:7–16. https://doi.org/10.1016/j.vetpar.2008.10.009
Ruiz DEM, Sempere LP, Martinez AG et al (2000) Technical and allocative efficiency analysis for cattle fattening on Argentina pampas. Agric Syst 65:179–199. https://doi.org/10.1016/S0308-521X(00)00032-9
Acknowledgements
We gratefully acknowledge the technicians and colleagues from the Animal Health Group at INTA Balcarce.
Funding
This work was funded by Research Grants from Agencia Nacional de Promoción Científica y Tecnológica, FONCyT, (PICT 2014–323), Universidad Nacional de Mar del Plata and Instituto Nacional de Tecnología Agropecuaria (PNSA 1115053, Argentina).
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Conceived and designed the experiments: MAD, YPH, JRC, DPM. Performed the experiments: MAD, JRC, YPH, VS, DPM. Analyzed the data: MAD, JRC. Contributed reagents/materials/analysis tools: JRC, LMOM, YPH, DPM, GJC, ELM. Wrote and commented on the manuscript: MAD, YPH, JRC, DPM, LMOM, ELM, GJC.
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Table S1
Multilocus microsatellite genotyping database used for this study (DOCX 60 kb)
Table S2
Diagnostic tests performed on N. caninum-infected fetuses submitted between 2008 and 2019 (DOCX 44 kb)
Fig. S1
Principal coordinate analysis (PCoA) based on the mean genetic distance between N. caninum MLGs from different countries. Geographic origin is indicated by the color (see legend). The proportion of variation for coordinate 1 is 52.22% (x axis) and for coordinate 2 (y axis) is 20.91% (PNG 8893 kb)
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Dorsch, M.A., Regidor-Cerrillo, J., Ortega-Mora, L.M. et al. Microsatellite genotyping reveals extensive genetic diversity in bovine Neospora caninum from the humid Pampa region in Argentina. Parasitol Res 119, 4049–4059 (2020). https://doi.org/10.1007/s00436-020-06922-x
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DOI: https://doi.org/10.1007/s00436-020-06922-x