Molecular survey of Cryptosporidium spp. in calves from the state of Mato Grosso, Brazil

Thábata dos Anjos Pacheco, Felippe Danyel Cardoso Martins, Sayanne Luns Hatum de Almeida, Thiago Borges Fernandes Semedo, Michelle Igarashi Watanabe, Valéria Dutra, João Luis Garcia, Richard Campos Pacheco

Abstract


Cryptosporidium spp. is a protozoan that infects a wide range of vertebrate hosts; it has been reported to be the cause of severe illness or death in livestock worldwide, which leads to decreased performance and production losses, especially in young animals. This study investigated the presence of Cryptosporidium in calves from beef farms in the state of Mato Grosso, midwestern Brazil. For this purpose, fecal samples from 237 animals aged ? 45 days, raised in 20 rural properties were subjected to DNA extraction and nested polymerase chain reaction (nPCR) targeting 18S ribosomal RNA (18S rRNA) gene followed by sequencing. Additionally, positive samples, previously identified as Cryptosporidium parvum by sequencing and phylogenetic analyses based on 18S rRNA gene, were subsequently analyzed focusing the amplification and sequencing using nPCR of a fragment of the 60 kDa glycoprotein (gp60) gene. Of the 237 fecal samples analyzed by PCR (18S rRNA), 50 (21.1%) fecal samples were positive for Cryptosporidium spp., while 14 (70%) of the 20 properties had at least one positive animal. The following Cryptosporidium species were detected: C. bovis, C. parvum, and C. ryanae. Thereafter, two potentially zoonotic subtypes (IIaA15G2R1 and IIaA16G3R1) of C. parvum were identified based on gp60 gene sequences. This study resulted in the detection of subtype IIaA16G3R1 for the first time in South America and showed a wide distribution of the protozoan in beef farms in the studied area of the State.

Keywords


Calf; Cryptosporidium parvum; Feces; Zoonosis.

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References


Brook, E., Hart, C. A., French, N., & Christley, R. (2008). Prevalence and risk factors for Cryptosporidium spp. infection in young calves. Veterinary Parasitology, 152(1-2), 46-52. doi: 10.1016/j.vetpar.2007.12. 003

Cavalier-Smith, T. (2014). Gregarine site-heterogeneous 18S rDNA trees, revision of gregarine higher classification, and the evolutionary diversification of Sporozoa. European Journal of Protistology, 50(5), 472-495. doi: 10.1016/j.ejop.2014.07.002

Holsback, L., Lima, H. E., Vidotto, O., Silva, M. A. D., Patelli, T. H. C., & Martins, F. D. C. (2018). Cryptosporidium occurrence in ruminants from the North Pioneer mesoregion of Paraná, Brazil. Revista Brasileira de Parasitologia Veterinária, 27(2), 248-253. doi: 10.1590/S1984-296120180037

Kimura, M. (1980). A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution, 16(2), 111-120. doi: 10. 1007/BF01731581

McAllister, T. A., Olson, M. E., Fletch, A., Wetzstein, M., & Entz, T. (2005). Prevalence of Giardia and Cryptosporidium in beef cows in southern Ontario and in beef calves in southern British Columbia. Canadian Veterinary Journal, 46(1), 47-55. doi: 10.4141/cjas66-008

Oliveira, J. P., F., Silva, D. P. G., Pacheco, M. D., Mascarini, L. M., Ribeiro, M. G., Alfieri, A. A. Borges, A.,... S. (2007). Diarréia em bezerros da raça Nelore criados extensivamente: estudo clínico e etiológico. Pesquisa Veterinária Brasileira, 27(10), 419-424. doi: 10.1590/S0100-736X2007001000006

Razakandrainibe, R., Diawara, E. H. I., Costa, D., Le Goff, L., Lemeteil, D., Ballet, J. J.,... Favennec, L. (2018). Common occurrence of Cryptosporidium hominis in asymptomatic and symptomatic calves in France. PLoS Neglected Tropical Diseases, 12(3), e0006355. doi: 10.1371/journal.pntd.0006355

Rieux, A., Paraud, C., Pors, I., & Chartier, C. (2014). Molecular characterization of Cryptosporidium isolates from beef calves under one month of age over three successive years in one herd in western France. Veterinary Parasitology, 202(3-4), 171-179. doi: 10.1016/j.vetpar.2014.03.004

Robertson, L. J., Björkman, C., Axén, C., & Fayer, R. (2014). Cryptosporidium: parasite and disease. In S. M. Caccio, & G. Widmer (Eds.), Cryptosporidium: parasite and disease (pp. 149-235). Wien: Springer-Verlag.

Silverlås, C., Bosaeus-Reineck, H., Näslund, K., & Björkman, C. (2013). Is there a need for improved Cryptosporidium diagnostics in Swedish calves? International Journal of Parasitology, 43(2), 155-161. doi: 10.1016/j.ijpara.2012.10.009

Silverlås, C., de Verdier, K., Emanuelson, U., Mattsson, J. G., & Björkman, C. (2010). Cryptosporidium infection in herds with and without calf diarrheal problems. Parasitology Research, 107(6), 1435-1444. doi: 10.1007/s00436-010-2020-x

Tamura, K., Stecher, G., Peterson, D., Filipski, A., & Kumar, S. (2013). MEGA6: molecular evolution-ary genetics analysis version 6.0. Molecular Biology and Evolution, 30(12), 2725-2729. doi: 10.1093/ molbev/mst197

Toledo, R. D., Martins, F. D., Ferreira, F. P., Almeida, J. C., Ogawa, L., Santos, H. L. dos,... Freire, R. L. (2017). Cryptosporidium spp. and Giardia spp. in feces and water and the associated exposure factors on dairy farms. PLoS One, 12(4), e0175311. doi: 10.1371/journal.pone.0175311

Trotz-Williams, L. A., Martin, D. S., Gatei, W., Cama, V., Peregrine, A. S., Martin, S. W.,... Xiao, L. (2006). Genotype and subtype analyses of Cryptosporidium isolates from dairy calves and humans in Ontario. Parasitology Research, 99(4), 346-352. doi: 10.1007/s00436-006-0157-4

Xiao, L. (2010). Molecular epidemiology of cryptosporidiosis: an update. Experimental Parasitology, 124(1), 80-89. doi: 10.1016/j.exppara.2009.03.018

Xiao, L., Morgan, U. M., Limor, J., Escalante, A., Arrowood, M., Shulaw, W.,… Lal, A. A. (1999). Genetic diversity within Cryptosporidium parvum and related Cryptosporidium species. Applied Environment Microbiology, 65(8), 3386-3391.




DOI: http://dx.doi.org/10.5433/1679-0359.2020v41n5supl1p2437

Semina: Ciênc. Agrár.
Londrina - PR
E-ISSN 1679-0359
DOI: 10.5433/1679-0359
E-mail: semina.agrarias@uel.br
Este obra está licenciado com uma Licença Creative Commons Atribuição-NãoComercial 4.0 Internacional