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Genetic diversity of Cryptosporidium spp. including novel identification of the Cryptosporidium muris and Cryptosporidium tyzzeri in horses in the Czech Republic and Poland

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Abstract

Faecal samples were collected from 352 horses on 23 farms operating under six different management systems in the Czech Republic and Poland during 2011 and 2012. Farms were selected without previous knowledge of parasitological status. All faecal samples were screened for Cryptosporidium spp. presence using microscopy, following aniline-carbol-methyl violet staining and PCR analysis of the small-subunit (SSU) rRNA and the 60-kDa glycoprotein (gp60) genes. Cryptosporidium muris-positive samples were additionally genotyped at four minisatellite markers: MS1 (encoding a hypothetical protein), MS2 (encoding a 90-kDa heat shock protein), MS3 (encoding a hypothetical protein) and MS16 (encoding a leucine-rich repeat family protein). Cryptosporidium spp. was detected by PCR in 12/352 (3.4 %) samples from 4 out of 13 farms. None of the samples tested by microscopy was positive. There was no relationship between Cryptosporidium prevalence and age, sex, diarrhoea or management system; however, Cryptosporidium was found only on farms where horses were kept on pasture during the day and in a stable overnight. Sequence analyses of SSU and gp60 genes revealed the presence of C. muris RN66 (n = 9), Cryptosporidium parvum IIaA15G2R1 (n = 1), Cryptosporidium tyzzeri IXbA22R9 (n = 1), and Cryptosporidium horse genotype VIaA15G4 (n = 1). The C. muris subtypes were identified as MS1-M1, MS2-M4, novel MS2-M7 and MS16-M1 by multilocus sequence of three minisatellite loci. The MS3 locus was not amplified from any isolate. This is the first report of C. tyzzeri and C. muris subtypes from horses.

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Acknowledgments

The authors would like to thank the farmers for their participation. This study was funded by the Grant of the Czech Science Foundation (15-01090S) and project of the agency of University of South Bohemia (011/2013/Z).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Faculty of Agriculture, University of South Bohemia in České Budějovice, Studentská 13, 370 05, České Budějovice, Czech Republic

    Pavla Wagnerová & Martin Kváč

  2. Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, v.v.i., Branišovská 31, 370 05, České Budějovice, Czech Republic

    Pavla Wagnerová, Bohumil Sak & Martin Kváč

  3. Department of Veterinary and Microbiological Sciences, North Dakota State University, Fargo, ND, USA

    John McEvoy

  4. Faculty of Economics, University of South Bohemia in České Budějovice, České Budějovice, Czech Republic

    Michael Rost

  5. Department of Parasitology, Institute of Genetics and Microbiology, Wroclaw University, Wroclaw, Poland

    Agniezska Perec Matysiak

  6. Faculty of Science, University of South Bohemia in České Budějovice, Studentská 13, 370 05, České Budějovice, Czech Republic

    Jana Ježková

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  1. Pavla Wagnerová
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  2. Bohumil Sak
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  4. Michael Rost
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  5. Agniezska Perec Matysiak
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  7. Martin Kváč
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Correspondence to Martin Kváč.

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Wagnerová, P., Sak, B., McEvoy, J. et al. Genetic diversity of Cryptosporidium spp. including novel identification of the Cryptosporidium muris and Cryptosporidium tyzzeri in horses in the Czech Republic and Poland. Parasitol Res 114, 1619–1624 (2015). https://doi.org/10.1007/s00436-015-4353-y

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  • DOI: https://doi.org/10.1007/s00436-015-4353-y

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