Cryptosporidium species detected in calves and cattle in Dagoretti, Nairobi, Kenya

Abstract

A total of 1,734 cattle faecal samples from 296 dairy-keeping households were collected from urban settings in Nairobi, Kenya. Modified Ziehl–Neelsen staining method and an immunofluorescence assay were used to identify those samples with Cryptosporidium oocyst infection. Oocysts from positive faecal samples were isolated by Sheather's sucrose flotation method and picked from the concentrate using cover slips. Genomic DNA was extracted from 124 of the faecal samples that were positive for Cryptosporidium and was used as template for nested PCR of the 18S rRNA gene. Twenty-five samples (20 %) were PCR-positive for Cryptosporidium, and 24 of the PCR products were successfully cloned and sequenced. Sequence and phylogenetic analysis identified 17 samples (68 %) as Cryptosporidium parvum-like, four samples (16 %) as Cryptosporidium ryanae, three samples (12 %) as Cryptosporidium andersoni and one sample (4 %) as Cryptosporidium hominis. To the best of our knowledge, this is the first genotyping study to report C. parvum-like, C. andersoni and C. hominis in cattle from Kenya. The results of this study show Cryptosporidium infections in calves and cattle may be potential zoonotic reservoirs of the parasite that infects humans.

This is a preview of subscription content, access via your institution.

Fig. 1

References

  1. Abreu-Acosta, N., Quispe, M.A., Foronda-Rodríguez, P., Alcoba-Florez, J., Lorenzo-Morales, J., Ortega-Rivas, A. and Valladares, B., 2007. Cryptosporidium in patients with diarrhoea, on Tenerife, Canary Islands, Spain, Annals of Tropical Medicine and Parasitology, 101, 539-545.

    PubMed  Article  CAS  Google Scholar 

  2. Amer, S., Honma, H., Ikarashi, M., Oishi, R., Endo, M., Otawa, K. and Nakai, Y., 2009. The first detection of Cryptosporidium deer-like genotype in cattle in Japan, Parasitology Research, 104, 745-752.

    PubMed  Article  Google Scholar 

  3. Brook, E., Hart, C.A., French, N.P. and Christley, R.P., 2008. Prevalence and risk factors for Cryptosporidium spp. Infection in young calves, Veterinary Parasitology, 152, 46–52.

    PubMed  Article  Google Scholar 

  4. Cacciò, S.M. and Pozio, E., 2006. Advances in the epidemiology, diagnosis and treatment of cryptosporidiosis, Expert Review of Anti-Infective.Therapy. 4, 429–443.

    Google Scholar 

  5. Casemore, D.P., Armstrong, M. and Sands, R.L., 1985. Laboratory diagnosis of cryptosporidiosis, Journal of Clinical Pathology, 38, 1337–1341.

    PubMed  Article  CAS  Google Scholar 

  6. Castro-Hermida, J.A., Almeida, A., González-Warleta, M., Correia da Costa, J.M., Rumbo-Lorenzo, C. and Mezo, M., 2007. Occurrence of Cryptosporidium parvum and Giardia duodenalis in healthy adult domestic ruminants, Parasitology Research, 101, 1443-1448.

    PubMed  Article  Google Scholar 

  7. Fayer, R., Santín, M. and Trout, JM., 2008. Cryptosporidium ryanae n. sp. Apicomplexa: Cryptosporidiidae in cattle Bos taurus, Veterinary Parasitology,156, 191–198.

    PubMed  Article  Google Scholar 

  8. Fayer, R., Santín, M. and Dargatz, D., 2010. Species of Cryptosporidium detected in weaned cattle on cow-calf operations in the United States, Veterinary Parasitology,170,187-192.

    PubMed  Article  Google Scholar 

  9. Fayer, R., Santín, M. and Trout, J.M., 2007. Prevalence of Cryptosporidium species and genotypes in mature dairy cattle on farms in eastern United States compared with younger cattle from the same locations, Veterinary Parasitology,145, 260-266.

    PubMed  Article  Google Scholar 

  10. Fayer, R., Santín, M., Trout, J.M. and Greiner, E., 2006. Prevalence of species and genotypes of Cryptosporidium found in 1-2-year-old dairy cattle in the eastern United States, Veterinary Parasitology, 135, 105-112.

    PubMed  Article  Google Scholar 

  11. Fayer, R., Santín, M. and Xiao, L., 2005. Cryptosporidium bovis n. sp. Apicomplexa, Cryptosporidiidae in cattle Bos taurus, Journal of Parasitology, 91, 624-629.

    PubMed  Article  Google Scholar 

  12. Felsenstein J. (1985). Confidence limits on phylogenies: An approach using the bootstrap. Evolution 39:783-791.

    Article  Google Scholar 

  13. Feltus, D.C., Giddings, C.W., Khaitsa, M.L. and McEvoy, J.M., 2008. High prevalence of Cryptosporidium bovis and the deer-like genotype in calves compared to mature cows in beef cow-calf operations, Veterinary Parasitology, 151, 191–195.

    PubMed  Article  Google Scholar 

  14. Feng, Y., Alderisio, K.A., Yang, W., Blancero, L.A., Kuhne, W.G., Nadareski, C.A., Reid, M. and Xiao, L., 2007. Cryptosporidium genotypes in wildlife from a new york watershed, Applied Environmental Microbiology, 73, 6475-6483.

    Article  CAS  Google Scholar 

  15. Gatei, W., Ashford, R.W., Beeching, N.J., Kamwati, S.K., Greensill, J. and Hart, C.A., 2002. Cryptosporidium muris infection in an HIV-infected adult, Kenya, Emerging Infectious Diseases, 8, 204-206.

    PubMed  Article  Google Scholar 

  16. Gatei, W., Greensill, J., Ashford, R.W., Cuevas, L.E., Parry, C.M., Cunliffe, N.A., Beeching, N.J. and Hart, C.A., 2003. Molecular analysis of the 18S rRNA gene of Cryptosporidium parasites from patients with or without human immunodeficiency virus infections living in Kenya, Malawi, Brazil, the United Kingdom, and Vietnam, Journal of Clinical Microbiology, 41, 1458-1462.

    PubMed  Article  CAS  Google Scholar 

  17. Gatei, W., Wamae, C.N., Mbae, C., Waruru, A., Mulinge, E., Waithera, T., Gatika, S.M., Kamwati, S.K., Revathi, G. and Hart, C.A., 2006. Cryptosporidiosis, prevalence, genotype analysis, and symptoms associated with infections in children in Kenya, American Journal of Tropical Medicine and Hygiene, 75, 78-82.

    PubMed  Google Scholar 

  18. Gibbons-Matthews, C., Prescott, A.M., 2003. Intra-isolate variation of Cryptosporidium parvum small subunit ribosomal RNA genes from human hosts in England, Parasitology Research, 90, 439-444.

    PubMed  Article  Google Scholar 

  19. Giles, M., Webster, K.A., Marshall, J.A., Catchpole, J. and Goddard, T.M., 2001. Experimental infection of a lamb with Cryptosporidium parvum genotype 1, Veterinary Record, 149, 523-525.

    PubMed  Article  CAS  Google Scholar 

  20. Halim, N.A., Plutzer, J., Bakheit, M.A. and Karanis, P., 2008. First report of Cryptosporidium deer-like genotype in Malaysian cattle, Veterinary Parasitology, 152, 325–329.

    PubMed  Article  CAS  Google Scholar 

  21. Jenkins, M.W., Tiwari, S., Lorente, M., Gichaba, C.M. and Wuertz, S., 2009. Identifying human and livestock sources of faecal contamination in Kenya with host-specific Bacteroidales assays, Water Research, 43, 4956-4966

    PubMed  Article  CAS  Google Scholar 

  22. Kang'ethe, E.K., McDermott, B., M'Ibui, G.M., Randolph, T.F., Langat, A.K., 2007. Investigation into the prevalence of bovine cryptosporidiosis among small-holder dairy households in Dagoretti Division, Nairobi, Kenya, East African Medical Journal, 8411 Suppl, S76-82.

    Google Scholar 

  23. Kang’ethe, E.K., Randolph, T.F., McDermott, B., Lang’ata, A.K., Kimani, V.N., Kirangu, M., Ekuttan, C., Ojigo, D., Onono, J., Ngonde, A. and Miriti, G., 2005. Characterization of benefits and health risks associated with smallholder dairy production in Dagoretti, Nairobi, Kenya. Final Technical Report submitted to the International Development Research Centre, Canada. IDRC, 102019-004.

  24. Keshavarz, A., Haghighi, A., Athari, A., Kazemi, B., Abadi, A. and Mojarad, E.N., 2009. Prevalence and molecular characterization of bovine Cryptosporidium in Qazvin province, Iran, Veterinary Parasitology, 160, 316-318.

    Article  CAS  Google Scholar 

  25. Khan, S.M., Debnath, C., Pramanik, A.K., Xiao, L., Nozaki, T. and Ganguly, S., 2010. Molecular characterization and assessment of zoonotic transmission of Cryptosporidium from dairy cattle in West Bengal, India, Veterinary Parasitology, 171, 41-47

    Article  CAS  Google Scholar 

  26. Kváč. M., Hromadová, N., Květoňová, D., Rost, M. and Sak, B., 2011. Molecular characterization of Cryptosporidium spp. in pre-weaned dairy calves in the Czech Republic: absence of C. ryanae and management-associated distribution of C. andersoni, C. bovis and C. parvum subtypes, Veterinary Parasitology, 177, 378-82.

    PubMed  Article  Google Scholar 

  27. Langkjaer, R.B., Vigre, H., Enemark, H.L. and Maddox-Hyttel, C., 2007. Molecular and phylogenetic characterization of Cryptosporidium and Giardia from pigs and cattle in Denmark. Parasitology 134, 339–350.

    PubMed  Article  CAS  Google Scholar 

  28. Liu, A., Wang, R., Li, Y., Zhang, L., Shu, J., Zhang, W., Feng, Y., Xiao , L. and Ling, H., 2009. Prevalence and distribution of Cryptosporidium spp. in dairy cattle in Heilongjiang Province, China, Parasitology Research. 105, 797-802.

    PubMed  Article  Google Scholar 

  29. Ma, P. and Soave, R., 1983. Three-step stool examination for cryptosporidiosis in 10 homosexual men with protracted watery diarrhea, Journal of Infectious Disease, 147, 824-828.

    Article  CAS  Google Scholar 

  30. Maikai, B.V., Umoh, J.U., Kwaga, J.K., Lawal, I.A., Maikai, V.A., Cama, V. and Xiao, L., 2011. Molecular characterization of Cryptosporidium spp. in native breeds of cattle in Kaduna State, Nigeria, Veterinary Parasitology. 178, 241-245.

    PubMed  Article  CAS  Google Scholar 

  31. Meireles, M.V., Soares, R.M., Bonello, F. and Gennari, S.M., 2007. Natural infection with zoonotic subtype of Cryptosporidium parvum in Capybara Hydrochoerus hydrochaeris from Brazil, Veterinary Parasitology, 147, 166-170.

    PubMed  Article  Google Scholar 

  32. Muchiri, J.M., Ascolillo, L., Mugambi, M., Mutwiri, T., Ward, H.D., Naumova, E.N., Egorov, A.I., Cohen, S., Else, J.G. and Griffiths, J.K., 2009. Seasonality of Cryptosporidium oocyst detection in surface waters of Meru, Kenya as determined by two isolation methods followed by PCR, Journal of Water and Health, 7, 67-75.

    PubMed  Article  CAS  Google Scholar 

  33. Nei, M. and Kumar, S., 2000, Molecular Evolution and Phylogenetics. Oxford University Press, New York.

    Google Scholar 

  34. Nydam, D.V., Wade, S.E., Schaaf, S.L. and Mohammed, H.O., 2001. Number of Cryptosporidium parvum oocysts or Giardia spp cysts shed by dairy calves after natural infection, American Journal of Veterinary Research, 62, 1612-1615.

    PubMed  Article  CAS  Google Scholar 

  35. Ondrácková, Z., Kvác, M., Sak, B., Kvetonová, D. and Rost, M., 2009. Prevalence and molecular characterization of Cryptosporidium spp. in dairy cattle in South Bohemia, the Czech Republic, Veterinary Parasitology, 165, 141-144.

    PubMed  Article  Google Scholar 

  36. Paul, S., Chandra, D., Tewari, A.K., Banerjee, P.S., Ray, D.D., Raina, O.K. and Rao, J.R., 2009. Prevalence of Cryptosporidium andersoni, a molecular epidemiological survey among cattle in India, Veterinary Parasitology, 161, 31-35.

    PubMed  Article  CAS  Google Scholar 

  37. Plutzer, J. and Karanis, P., 2007. Genotype and subtype analyses of Cryptosporidium isolates from cattle in Hungary, Veterinary Parasitology, 146, 357–362.

    PubMed  Article  CAS  Google Scholar 

  38. Ryan, U.M., Bath, C., Robertson, I., Read, C., Elliot, A., McInnes, L., Traub, R. and Besier, B., 2005. Sheep may not be an important zoonotic reservoir for Cryptosporidium and Giardia parasites, Applied Environmental Microbiology, 71, 4992-4997.

    Article  CAS  Google Scholar 

  39. Ryan, U.M., Samarasinghe, B., Read, C., Buddle, J.R., Robertson, I.D. and Thompson, R.C.A., 2003. Identification of a novel Cryptosporidium genotype in pigs, Applied Environmental Microbiology, 69, 3970–3974.

    Article  CAS  Google Scholar 

  40. Saitou, N. and Nei, M., 1987. The neighbour -joining method: Anew method for reconstructing phylogenetic trees. Molecular Biology and Evolution,4, 406-425.

    Google Scholar 

  41. Santı´n, M., Trout, J.M. and Fayer, R., 2008. A longitudinal study of cryptosporidiosis in dairy cattle from birth to two years of age, Veterinary Parasitology,155, 15-23.

    PubMed  Article  Google Scholar 

  42. Santín, M., Trout, J.M., Xiao, L., Zhou, L., Greiner, E. and Fayer, R., 2004. Prevalence and age-related variation of Cryptosporidium species and genotypes in dairy calves, Veterinary Parasitology, 122, 103–117.

    PubMed  Article  Google Scholar 

  43. Silverlås, C., Emanuelson, U., de Verdier, K. and Björkman, C., 2009. Prevalence and associated management factors of Cryptosporidium shedding in 50 Swedish dairy herds, Preventive Veterinary Medicine, 90, 242-253.

    PubMed  Article  Google Scholar 

  44. Siwila, J., Phiri, I.G., Vercruysse, J., Goma, F., Gabriel, S., Claerebout, E., and Geurden, T., 2007. Asymptomatic cryptosporidiosis in Zambian dairy farm workers and their household members, Transactions of the Royal Society for Tropical Medicine and Hygiene, 101, 733–734.

    Article  Google Scholar 

  45. Smith, H.V., Nichols, R.A., Mallon, M., Macleod, A., Tait, A., Reilly, W.J., Browning, L.M., Gray, D., Reid, S.W. and Wastling, J.M., 2005 Natural Cryptosporidium hominis infections in Scottish cattle, Veterinary Record, 156, 710–711.

    PubMed  CAS  Google Scholar 

  46. Soba, B., Petrovec, M., Mioc, V. and Logar, J., 2006. Molecular characterisation of Cryptosporidium isolates from humans in Slovenia, Clinical Microbiology and Infection, 12, 918-921.

    PubMed  Article  CAS  Google Scholar 

  47. Soltane, R., Guyot, K., Dei-Cas, E. and Ayadi, A., 2007. Cryptosporidium parvum Eucoccidiorida, Cryptosporiidae in calves, results of a longitudinal study in a dairy farm in Sfax, Tunisia, Parasitology, 14, 309-312.

    CAS  Google Scholar 

  48. Szonyi, B., Kang’ethe, E.K., Mbae, C.K., Kakundi, E.M., Kamwati, S.K. and Mohammed, H.O., 2008. First report of Cryptosporidium deer-like genotype in Kenyan cattle, Veterinary Parasitology, 153, 172–175.

    PubMed  Article  Google Scholar 

  49. Tamura K., Peterson D., Peterson N., Stecher G., Nei M., and Kumar S., 2011. MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Molecular Biology and Evolution doi:10.1093/molbev/msr121.

  50. Thomaz, A., Meireles, M.V., Soares, R.M., Pena, H.F. and Gennari, S.M., 2007. Molecular identification of Cryptosporidium spp. from faecal samples of felines, canines and bovines in the state of São Paulo, Brazil, Veterinary Parasitology,150, 291-296.

    PubMed  Article  CAS  Google Scholar 

  51. Thompson, H.P., Dooley, J.S., Kenny, J., McCoy, M., Lowery, C.J., Moore, J.E., and Xiao, L., 2007. Genotypes and subtypes of Cryptosporidium spp. in neonatal calves in Northern Ireland, Parasitology Research, 100, 619-624

    PubMed  Article  Google Scholar 

  52. Thornton, C.G. and Passen, S., 2004. Inhibition of PCR amplification by phytic acid, and treatment of bovine faecal specimens with phytase to reduce inhibition, Journal of Microbiological Methods, 59, 43–52.

    PubMed  Article  CAS  Google Scholar 

  53. Wielinga, P.R., de Vries, A., van der Goot, T.H., Mank, T., Mars, M.H., Kortbeek, L.M. and van der Giessen, J.W., 2008. Molecular epidemiology of Cryptosporidium in humans and cattle in The Netherlands, International Journal of Parasitology, 38, 809-817

    PubMed  Article  CAS  Google Scholar 

  54. Xiao, L. and Ryan, U.M., 2004. Cryptosporidiosis, an update in molecular epidemiology, Current Opinion in Infectious Disease, 17, 483-490.

    Article  Google Scholar 

Download references

Acknowledgements

The authors wish to thank the International Research Development Centre for funding this work under the grant 103075-001. We also thank all the members of the research team involved in this project for the support and encouragement in various ways.

We also acknowledge the support given by the BecA-ILRI Hub, for their excellent technical support.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Erastus K. Kang’ethe.

Additional information

This paper is part of a special supplement on assessing and managing urban zoonoses and food-borne disease in two African cities (Nairobi, Kenya and Ibadan, Nigeria).

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kang’ethe, E.K., Mulinge, E.K., Skilton, R.A. et al. Cryptosporidium species detected in calves and cattle in Dagoretti, Nairobi, Kenya. Trop Anim Health Prod 44, 25–31 (2012). https://doi.org/10.1007/s11250-012-0202-5

Download citation

Keywords

  • Cryptosporidium
  • Genotyping
  • Dairy farming
  • Kenya