Detection and molecular characterization of Anaplasma marginale in DNA from blood samples of bovines from Paraguay
DOI:
https://doi.org/10.18004/Keywords:
DNA, Anaplasma marginale, cPCR, bovine, bloodAbstract
Anaplasma marginale is an intraerythrocytic bacteria responsible for the disease known as bovine anaplasmosis, which has a great impact on livestock in tropical and subtropical regions. The importance of using conventional PCR (cPCR) is the early detection of persistent infections, since other study methods may present limitations when are diagnosed. The region of the Msp5 gene is specific for the detection of persistent infections. The present work was carried out using 86 bovine DNA samples, from whole blood using the cPCR technique for the amplification of the Msp5 gene. Thirty five percent (30/86) of the samples were positive for Anaplasma marginale. Of the positive samples, three were sequenced. This analysis determined the presence of A. marginale with an identity level of 99 – 100%. The construction of the phylogenetic tree showed that the samples obtained were closely related to other sequences previously reported in countries from Asia and Africa. This study shows prevalence of the disease in Paraguayan cattle. Furthermore, it is noted that this is the first molecular evidence of A. marginale in Paraguay done by molecular techniques, so it is recommended to expand studies on its pathogenicity, transmission mechanism and geographical distribution.
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Altschul, S. F., Gish, W., Miller, W., Myers, E. W., & Lipman, D. J. (1990). Basic local alignment search tool. Journal of Molecular Biology, 215(3), 403-410. https://doi.org/10.1016/S0022-2836(05)80360-2
Arnuphapprasert, A., Nugraheni, Y. R., Poofery, J., Aung, A., Kaewlamun, W., Chankeaw, W., Tasanaganjanakorn, T., Wattanamethanont, J., & Kaewthamasorn, M. (2023). Genetic characterization of genes encoding the major surface proteins of Anaplasma marginale from cattle isolates in Thailand reveals multiple novel variants. Ticks and tick-borne diseases, 14(2), 102110. https://doi.org/10.1016/j.ttbdis.2022.102110
Aubry, P., & Geale, D. W. (2010). A review of bovine anaplasmosis. Transboundary and emerging diseases, 58(1), 1–30. https://doi.org/10.1111/j.1865-1682.2010.01173.x
Balbuena, E. A. (2021). Frecuencia de Anaplasmosis subclínica en bovinos adultos en el tambo lechero de la Facultad de Ciencias Veterinarias, Universidad Nacional de Asunción en el año 2021 (Trabajo de grado). Facultad de Ciencias Veterinarias, Universidad Nacional de Asunción. 74 pag.
Barbet, A. F., Lundgren, A., Yi, J., Rurangirwa, F. R., & Palmer, G. H. (2000). Antigenic variation of Anaplasma marginale by expression of MSP2 mosaics. Infection and immunity, 68(11), 6133–6138. https://doi.org/10.1128/IAI.68.11.6133-6138.2000
Ben Said, M., Belkahia, H., & Messadi, L. (2018). Anaplasma spp. in North Africa: A review on molecular epidemiology, associated risk factors and genetic characteristics. Ticks and tick-borne diseases, 9(3), 543–555. https://doi.org/10.1016/j.ttbdis.2018.01.003
Brayton, K. A., Kappmeyer, L. S., Herndon, D. R., Dark, M. J., Tibbals, D. L., Palmer, G. H., McGuire, T. C., & Knowles, D. P. (2005). Complete genome sequencing of Anaplasma marginale reveals that the surface is skewed to two superfamilies of outer membrane proteins. Proceedings of the National Academy of Sciences of the United States of America, 102(3), 844–849. https://doi.org/10.1073/pnas.0406656102
Brito, L. G., Sena Oliveira, M. C. de, Fonseca Moura, M. N. da, da Silva Netto, F. G., Cavalcante, F. A., Marim, A. D., Rodrigues de Souza, G. C., & da Silva, J. L. (2006). Extração de DNA a partir de coágulos sanguíneos bovinos. Porto Velho. Embrapa. https://ainfo.cnptia.embrapa.br/digital/bitstream/item/24667/1/bpd43-dnabovinos.pdf
Camacho-Nuez, M., de Lourdes Muñoz, M., Suarez, C. E., McGuire, T. C., Brown, W. C., & Palmer, G. H. (2000). Expression of polymorphic msp1beta genes during acute Anaplasma marginale rickettsemia. Infection and immunity, 68(4), 1946–1952. https://doi.org/10.1128/IAI.68.4.1946-1952.2000
Canever, M. F., Vieira, L. L., Reck, C., Richter, L., & Miletti, L. C. (2014). First evaluation of an outbreak of bovine babesiosis and anaplasmosis in Southern Brazil using multiplex PCR. The Korean journal of parasitology, 52(5), 507–511. https://doi.org/10.3347/kjp.2014.52.5.507
Castro, L. M. (2019). Frecuencia de Anaplasma spp. y Babesia spp. en bovinos de carne en un establecimiento ganadero en la ciudad de Caacupu, departamento de Paraguari en el año 2019 (Trabajo de grado). Facultad de Ciencias Veterinarias, Universidad Nacional de Asunción. 78 pag.
Corona, B. & Martínez, S. (2011). Deteccción de Anaplasma marginale en bovinos, mediante la amplificación por PCR del gen msp5. Revista de Salud Animal, 33(1), 24-31. http://scielo.sld.cu/scielo.php?script=sci_arttext&pid=S0253-570X2011000100004&lng=es&tlng=es
da Silva, J. B., André, M. R., & Machado, R. Z. (2016). Low genetic diversity of Anaplasma marginale in calves in an endemic area for bovine anaplasmosis in the state of São Paulo, Brazil. Ticks and tick-borne diseases, 7(1), 20–25. https://doi.org/10.1016/j.ttbdis.2015.07.018
de la Fuente, J., Lew, A., Lutz, H., Meli, M. L., Hofmann-Lehmann, R., Shkap, V., Molad, T., Mangold, A. J., Almazán, C., Naranjo, V., Gortázar, C., Torina, A., Caracappa, S., García-Pérez, A. L., Barral, M., Oporto, B., Ceci, L., Carelli, G., Blouin, E. F., & Kocan, K. M. (2005). Genetic diversity of Anaplasma species major surface proteins and implications for anaplasmosis serodiagnosis and vaccine development. Animal health research reviews, 6(1), 75–89. https://doi.org/10.1079/ahr2005104
de la Fuente, J., & Kocan, K. M. (2000). Anaplasma marginale: biology and control. In: Kocan K. M. (Ed.), Anaplasma marginale. Dordrecht: Springer
de Souza Ramos, I., Herrera, H. M., Fernandes, S. J., do Amaral, R. B., de Souza Zanatto, D. C., Viera da Silva, T., Le Senechal Horta, B., Vilela Campos, J. B., Almeida Alves, J. V., Carvalho de Macedo, G., Zacarias Machado, R., & Rogério André, M. (2019). Genetic diversity of Anaplasma marginale in beef cattle in the Brazilian Pantanal. Ticks and Tick-borne Diseases, 10(4), 805-814. https://doi.org/10.1016/j.ttbdis.2019.03.015
Dumler, J. S., Barbet, A. F., Bekker, C. P., Dasch, G. A., Palmer, G. H., Ray, S. C., Rikihisa, Y., & Rurangirwa, F. R. (2001). Reorganization of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and 'HGE agent' as subjective synonyms of Ehrlichia phagocytophila. International journal of systematic and evolutionary microbiology, 51(6), 2145–2165. https://doi.org/10.1099/00207713-51-6-2145
Espinoza, A. J. (2018). Determinacion de Anaplasmosis y Leucosis viral bovina en dos tambos del departamento central (San Lorenzo y Ñemby) en el año 2018 (Trabajo de grado). Facultad de Ciencias Veterinarias, Universidad Nacional de Asunción. 60 pag.
French, D. M., McElwain, T. F., McGuire, T. C., & Palmer, G. H. (1998). Expression of Anaplasma marginale major surface protein 2 variants during persistent cyclic rickettsemia. Infection and immunity, 66(3), 1200–1207. https://doi.org/10.1128/IAI.66.3.1200-1207.1998
Futse, J. E., Ueti, M. W., Knowles, D. P., Jr, & Palmer, G. H. (2003). Transmission of Anaplasma marginale by Boophilus microplus: retention of vector competence in the absence of vector-pathogen interaction. Journal of clinical microbiology, 41(8), 3829–3834. https://doi.org/10.1128/JCM.41.8.3829-3834.2003
Goff, W. L., McElwain, T. F., Suarez, C. E., Johnson, W. C., Brown, W. C., Norimine, J., & Knowles, D. P. (2003). Competitive enzyme-linked immunosorbent assay based on a rhoptry-associated protein 1 epitope specifically identifies Babesia bovis-infected cattle. Clinical and diagnostic laboratory immunology, 10(1), 38–43. https://doi.org/10.1128/cdli.10.1.38-43.2003
Herrero, M. V., Pérez, E., Goff, W. L., Torioni, E. S., Knowles, D. P., McElwain, T. F., Álvarez, V., Álvarez, A., & Buening, G. M. (1998). Herrero, M. V., Perez, E., Goff, W. L., Torioni de Echaide, S., Knowles, D. P., McElwain, T. F., Alvarez, V., Alvarez, A., & Buening, G. M. (1998). Prospective study for the detection of Anaplasma marginale Theiler, 1911 (Rickettsiales: Anaplasmataceae) in Costa Rica. Annals of the New York Academy of Sciences, 849, 226–233. https://doi.org/10.1111/j.1749-6632.1998.tb11053.x
Kocan, K. M., de la Fuente, J., Blouin, E. F., Coetzee, J. F., & Ewing, S. A. (2010). The natural history of Anaplasma marginale. Veterinary parasitology, 167(2-4), 95–107. https://doi.org/10.1016/j.vetpar.2009.09.012
Kocan, K. M., de la Fuente, J., Guglielmone, A. A., & Meléndez, R. D. (2003). Antigens and alternatives for control of Anaplasma marginale infection in cattle. Clinical microbiology reviews, 16(4), 698–712. https://doi.org/10.1128/CMR.16.4.698-712.2003
Kocan, K. M., de la Fuente, J., Blouin, E. F., & Garcia-Garcia, J. C. (2004). Anaplasma marginale (Rickettsiales: Anaplasmataceae): recent advances in defining host-pathogen adaptations of a tick-borne rickettsia. Parasitology, 129 Suppl, S285–S300. https://doi.org/10.1017/s0031182003004700
Kumar, S., Stecher, G., Li, M., Knyaz, C., & Tamura, K. (2018). MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. Molecular biology and evolution, 35(6), 1547–1549. https://doi.org/10.1093/molbev/msy096
Melman, S., Hartt, Y., & Giardina, S. (2004). El uso de la biología molecular en el diagnóstico de Anaplasma marginale. Croizatia, 3, 40-52.
Nava, S., Lareschi, M., Rebollo, C., Benítez Usher, C., Beati, L., Robbins, R. G., Durden, L. A., Mangold, A. J., & Guglielmone, A. A. (2007). The ticks (Acari: Ixodida: Argasidae, Ixodidae) of Paraguay. Annals of tropical medicine and parasitology, 101(3), 255–270. https://doi.org/10.1179/136485907X176319
Organización Mundial de Sanidad Animal (2024). Anaplasmosis bovina: manual de las pruebas de diagnostico y de las vacunas para los animales terrestres. Paris. OIE https://www.woah.org/es/que-hacemos/normas/codigos-y-manuales/
Ogata, S., Pereira, J. A. C., Jhonny, L. V. A., Carolina, H. P. G., Matsuno, K., Orba, Y., Sawa, H., Kawamori, F., Nonaka, N., & Nakao, R. (2021). Molecular Survey of Babesia and Anaplasma Infection in Cattle in Bolivia. Veterinary sciences, 8(9), 188. https://doi.org/10.3390/vetsci8090188
Palmer, G. H., Brown, W. C., & Rurangirwa, F. R. (2000). Antigenic variation in the persistence and transmission of the ehrlichia Anaplasma marginale. Microbes and infection, 2(2), 167–176. https://doi.org/10.1016/s1286-4579(00)00271-9
Parodi, P., Corbellini, L. G., Leotti, V. B., Rivero, R., Miraballes, C., Riet-Correa, F., Venzal, J. M., & Armúa-Fernández, M. T. (2021). Validation of a multiplex PCR assay to detect Babesia spp. and Anaplasma marginale in cattle in Uruguay in the absence of a gold standard test. Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc, 33(1), 73–79. https://doi.org/10.1177/1040638720975742
Rymaszewska, A., & Grenda, S. (2008). Bacteria of the genus Anaplasma - characteristics of Anaplasma and their vectors: a review. Veterinární medicína, 53(11), 573-584. doi: 10.17221/1861-VETMED
Torioni de Echaide, S., Knowles, D. P., McGuire, T. C., Palmer, G. H., Suarez, C. E., & McElwain, T. F. (1998). Detection of cattle naturally infected with Anaplasma marginale in a region of endemicity by nested PCR and a competitive enzyme-linked immunosorbent assay using recombinant major surface protein 5. Journal of clinical microbiology, 36(3), 777–782. https://doi.org/10.1128/JCM.36.3.777-782.1998
Tamekuni, K., Kano, F. S., Ataliba, A. C., Marana, E. R., Venâncio, E. J., Vidotto, M. C., Garcia, J. L., Headley, S. A., & Vidotto, O. (2009). Cloning, expression, and characterization of the MSP1a and MSP1b recombinant proteins from PR1 Anaplasma marginale strain, Brazil. Research in veterinary science, 86(1), 98–107. https://doi.org/10.1016/j.rvsc.2008.05.016
Wahba, A. (2017). Serological and microscopical diagnosis of anaplasmosis in some farm animals. Animal Health Research Journal, 5 (4), pp. 68–75. https://www.researchgate.net/
publication/323726100_Serological_and_microscopical_diagnosis_of_anaplasmosis_in_some_farm_animals
Zim, M. M. R., Ahmed, N., Ahmed, M., Miah, M. A. H., Sajib, M. M. R., Rabbi, M. R. R., Rahman, M. K., Roy, B. C., & Talukder, M. H. (2024). First seroprevalence survey of bovine anaplasmosis: an emerging tick-borne disease in commercial livestock and dairy farms in Bangladesh. Parasitology, 151(9), 992–1000. https://doi.org/10.1017/S0031182024001495.
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Copyright (c) 2025 Olga Lorena Nuñez Yegros, Maria de Lourdes Narvaez Quiñonez, Alba Maria Araujo Oviedo, Guillermo Giménez Bareiro, Ingrid Paloma Florentín Ibarra, Lino César Ramírez Ayala
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