Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter December 28, 2016

Report of fatal mixed infection with Cryptosporidium parvum and Giardia intestinalis in neonatal calves

Yuu Matsuura, Makoto Matsubayashi, Satoko Nukata, Tomoyuki Shibahara, Osamu Ayukawa, Yasuko Kondo, Tomohide Matsuo, Shigehiko Uni, Masaru Furuya, Hiroyuki Tani, Naotoshi Tsuji and Kazumi Sasai
From the journal Acta Parasitologica


In the production and management of beef and dairy cattle, controlling diarrhea is one of the important concerns. Pathogenic agents of the disease, protozoan parasites including Cryptosporidium spp., are difficult to control, making prevention, diagnoses, and treatment of diarrhea. In the present study, we investigated a farm with a history of calf deaths over a period of 10 years in order to determine the cause of disease and to clarify the detailed distribution of the pathogens. In four examined calves that were reared in calf pens, all were positive with Cryptosporidium and/or Giardia, while the other breeding stock and adult cattle were negative. Molecular analyses revealed that the isolates from calves were C. parvum subtype IIaA15G2R1 as a zoonotic and G. intestinalis assemblage E. Other pathogenic bacteria and diarrhea-causing viruses were not detected. After treating the calf pens with boiling water and milk of lime (Ca[OH]2), oocysts of C. parvum and cysts of G. intestinalis were not found and no additional calves died. This is the first report to describe the mixed infection of both parasites in Japan.


Abe N., Kimata I., Iseki M. 2002. Identification of genotypes of Cryptosporidium parvum isolates from a patient and a dog in Japan. Journal of Veterinary Medical Science, 64, 165–16810.1292/jvms.64.165Search in Google Scholar

Appelbee A.J., Frederick L.M., Heitman T.L., Olson M.E. 2003. Prevalence and genotyping of Giardia duodenalis from beef calves in Alberta, Canada. Veterinary Parasitology, 112, 289–29410.1016/S0304-4017(02)00422-3Search in Google Scholar

Bajer A. 2008. Cryptosporidium and Giardia spp. infections in humans, animals and the environment in Poland. Parasitology Research, 104, 1–1710.1007/s00436-008-1179-xSearch in Google Scholar

Ballweber L.R., Xiao L., Bowman D.D., Kahn G., Cama V.A. 2010. Giardiasis in dogs and cats: update on epidemiology and public health significance. Trends in Parasitology, 26, 180–18910.1016/ in Google Scholar

Barigye R., Dyer N.W., Newell T.K., Khaitsa M.L., Trout J.M., Santin M., Fayer R. 2008. Molecular and immunohistochemical detection of assemblage E, Giardia duodenalis in scouring North Dakota calves. Veterinary Parasitology, 157, 196–20210.1016/j.vetpar.2008.07.033Search in Google Scholar

Cacciò S.M., De Giacomo M., Pozio E. 2002. Sequence analysis of the beta-giardin gene and development of a polymerase chain reaction-restriction fragment length polymorphism assay to genotype Giardia duodenalis cysts from human faecal samples. International Journal for Parasitology, 32, 1023–103010.1016/S0020-7519(02)00068-1Search in Google Scholar

Carranza P.G., Lujan H.D. 2010. New insights regarding the biology of Giardia lamblia. Microbes and Infection, 12, 71–8010.1016/j.micinf.2009.09.008Search in Google Scholar PubMed

Chauret C.P., Radziminski C.Z., Lepuil M., Creason R., Andrews R.C. 2001. Chlorine dioxide inactivation of Cryptosporidium parvum oocysts and bacterial spore indicators. Applied and Environmental Microbiology, 67, 2993–300110.1128/AEM.67.7.2993-3001.2001Search in Google Scholar PubMed PubMed Central

Cho Y.I., Yoon K.J. 2014. An overview of calf diarrhea – infectious etiology, diagnosis, and intervention. Journal of Veterinary Science, 15, 1–1710.4142/jvs.2014.15.1.1Search in Google Scholar PubMed PubMed Central

De Cesaro M.P., Pierezan F., Heins B.D., Brown C.C. 2014. Pathology in practice.Bovine cryptosporidiosis. Journal of the American Veterinary Medical Association, 244, 53–5510.2460/javma.244.1.53Search in Google Scholar PubMed

Esch K.J., Petersen C.A. 2013. Transmission and epidemiology of zoonotic protozoal diseases of companion animals. Clinical Microbiology Reviews, 26, 58–8510.1128/CMR.00067-12Search in Google Scholar PubMed PubMed Central

Fujino T., Matsui T., Kobayashi F., Haruki K., Yoshino Y., Kajima J., Tsuji M. 2002.The effect of heating against Cryptosporidium oocysts. Journal of Veterinary Medical Science, 64, 199–20010.1292/jvms.64.199Search in Google Scholar PubMed

Fukuda M., Kuga K., Miyazaki A., Suzuki T., Tasei K., Aita T., Mase M., Sugiyama M., Tsunemitsu H. 2012. Development and application of one-step multiplex reverse transcription PCR for simultaneous detection of five diarrheal viruses in adult cattle. Archives of Virology, 157, 1063–106910.1007/s00705-012-1271-5Search in Google Scholar PubMed PubMed Central

Gillhuber J., Rügamer D., Pfister K., Scheuerle M.C. 2014. Giardiosis and other enteropathogenic infections: a study on diarrhoeic calves in Southern Germany. BMC Research Notes, 7, 11210.1186/1756-0500-7-112Search in Google Scholar PubMed PubMed Central

Graczyk T.K., Kacprzak M., Neczaj E., Tamang L., Graczyk H., Lucy F.E., Girouard A.S. 2008. Occurrence of Cryptosporidium and Giardia in sewage sludge and solid waste landfill leachate and quantitative comparative analysis of sanitization treatments on pathogen inactivation. Environmental Research, 106, 27–3310.1016/j.envres.2007.05.005Search in Google Scholar PubMed

Heyworth M.F. 2016. Giardia duodenalis genetic assemblages and hosts. Parasite, 23, 1310.1051/parasite/2016013Search in Google Scholar PubMed PubMed Central

Ichikawa-Seki M., Aita J., Masatani T., Suzuki M., Nitta Y., Tamayose G., Iso T., Suganuma K., Fujiwara T., Matsuyama K., Niikura T., Yokoyama N., Suzuki H., Yamakawa K., Inokuma H., Itagaki T., Zakimi S., Nishikawa Y. 2015.10.1016/j.parint.2014.11.007Search in Google Scholar PubMed PubMed Central

Itagaki T., Kinoshita S., Aoki M., Itoh N., Saeki H., Sato N., Uetsuki J., Izumiyama S., Yagita K., Endo T. 2005. Genotyping of Giardia intestinalis from domestic and wild animals in Japan using glutamete dehydrogenase gene sequencing. Veterinary Parasitology, 133, 283–28710.1016/j.vetpar.2005.05.061Search in Google Scholar PubMed

Koompapong K., Sutthikornchai C., Sukthana Y. 2009. Cryptosporidium oocyst detection in water samples: floatation technique enhanced with immunofluorescence is as effective as immunomagnetic separation method. Korean Journal of Parasitology, 47, 353–35710.3347/kjp.2009.47.4.353Search in Google Scholar PubMed PubMed Central

Liu A., Zhang X., Zhang L., Wang R., Li X., Shu J., Zhang X., Shen Y., Zhang W., Ling H. 2012. Occurrence of bovine giardiasis and endemic genetic characterization of Giardia duodenalis isolates in Heilongjiang Province, in the Northeast of China. Parasitology Research, 111, 655–66110.1007/s00436-012-2883-0Search in Google Scholar PubMed

Lorenz I., Fagan J., More S.J. 2011. Calf health from birth to weaning. II. Management of diarrhoea in pre-weaned calves. Irish Veterinary Journal, 64, 910.1186/2046-0481-64-9Search in Google Scholar

Matsubayashi M., Kimata I., Abe N. 2005. Identification of genotypes of Giardia intestinalis isolates from a human and calf in Japan. Journal of Veterinary Medical Science, 67, 337–34010.1292/jvms.67.337Search in Google Scholar

Matsubayashi M., Teramoto-Kimata I., Uni S., Lillehoj H.S., Matsuda H., Furuya M., Tani H., Sasai K. 2013. Elongation factor-1a is a novel protein associated with host cell invasion and a potential protective antigen of Cryptosporidium parvum. Journal of Biological Chemistry, 288, 34111–3412010.1074/jbc.M113.515544Search in Google Scholar

Meganck V., Hoflack G., Opsomer G. 2014. Advances in prevention and therapy of neonatal dairy calf diarrhoea: a systematical review with emphasis on colostrum management and fluid therapy. Acta Veterinaria Scandinavica, 56, 7510.1186/s13028-014-0075-xSearch in Google Scholar

Nagano S., Matsubayashi M., Kita T., Narushima T., Kimata I., Iseki M., Hajiri T., Tani H., Sasai K., Baba E. 2007. Detection of a mixed infection of a novel Cryptosporidium andersoni and its subgenotype in Japanese cattle. Veterinary Parasitology, 149, 213–21810.1016/j.vetpar.2007.07.016Search in Google Scholar

O’Handley R.M., Cockwill C., McAllister T.A., Jelinski M., Morck D.W., Olson M.E. 1999. Duration of naturally acquired giardiosis and cryptosporidiosis in dairy calves and their association with diarrhea. Journal of the American Veterinary Medical Association, 214, 391–396Search in Google Scholar

Peng M.M., Matos O., Gatei W., Das P., Stantic-Pavlinic M., Bern C., Sulaiman I.M., Glaberman S., Lal A.A., Xiao L. 2001.A comparison of Cryptosporidium subgenotypes from several geographic regions. Journal of Eukaryotic Microbiology, Suppl, 28S–31S10.1111/j.1550-7408.2001.tb00442.xSearch in Google Scholar

Ralston B.J., McAllister T.A., Olson M.E. 2003. Prevalence and infection pattern of naturally acquired giardiasis and cryptosporidiosis in range beef calves and their dams. Veterinary Parasitology, 114, 113–12210.1016/S0304-4017(03)00134-1Search in Google Scholar

Rimhanen-Finne R., Vuorinen A., Marmo S., Malmberg S., Hänninen M.L. 2004. Comparative analysis of Cryptosporidium, Giardia and indicator bacteria during sewage sludge hygienization in various composting processes. Letters in Applied Microbiology, 38, 301–30510.1111/j.1472-765X.2004.01487.xSearch in Google Scholar

Ryan U., Fayer R., Xiao L. 2014. Cryptosporidium species in humans and animals: current understanding and research needs. Parasitology, 141, 1667–168510.1017/S0031182014001085Search in Google Scholar PubMed

Robertson L.J., Campbell A.T., Smith H.V. 1992.Survival of Cryptosporidium parvum oocysts under various environmental pressures. Applied and Environmental Microbiology, 58, 3494–350010.1128/aem.58.11.3494-3500.1992Search in Google Scholar PubMed PubMed Central

Smith G. 2015. Antimicrobial decision making for enteric diseases of cattle. Veterinary Clinics of North America: Food Animal Practice, 31, 47–6010.1016/j.cvfa.2014.11.004Search in Google Scholar

Uga S., Matsuo J., Kono E., Kimura K., Inoue M., Rai S.K., Ono K. 2000. Prevalence of Cryptosporidium parvum infection and pattern of oocyst shedding in calves in Japan. Veterinary Parasitology, 94, 27–3210.1016/S0304-4017(00)00338-1Search in Google Scholar

Vilcek S. 1994. Development of PCR tests for the detection of bovine herpesvirus-1, bovine respiratory syncytial viruses and pestiviruses. Veterinární medicína, 39, 687–700Search in Google Scholar

Vu-Khac H., Holoda E., Pilipcinec E., Blanco M., Blanco J.E., Dahbi G., Mora A., López C., González E.A., Blanco J. 2007. Serotypes, virulence genes, intimin types and PFGE profiles of Escherichia coli isolated from piglets with diarrhoea in Slovakia. Veterinary Journal, 174, 176–18710.1016/j.tvjl.2006.05.019Search in Google Scholar PubMed

Xiao L. 2010. Molecular epidemiology of cryptosporidiosis: an update. Experimental Parasitology, 124, 80–8910.1016/j.exppara.2009.03.018Search in Google Scholar PubMed

Xiao L., Morgan U.M., Limor J., Escalante A., Arrowood M., Shulaw W., Thompson R.C., Fayer R., Lal A.A. 1999. Genetic diversity within Cryptosporidium parvum and related Cryptosporidium species. Applied and Environmental Microbiology, 65, 3386–339110.1128/AEM.65.8.3386-3391.1999Search in Google Scholar PubMed PubMed Central

Yui T., Nakajima T., Yamamoto N., Kon M., Abe N., Matsubayashi M., Shibahara T. 2014. Age-related detection and molecular characterization of Cryptosporidium suis and Cryptosporidium scrofarum in pre– and post-weaned piglets and adult pigs in Japan. Parasitology Research,113, 359–36510.1007/s00436-013-3662-2Search in Google Scholar PubMed

Zintl A., Keogh B., Ezzaty-Mirhashemi M., De Waal T., Scholz D., Mulcahy G. 2010.Survival of Cryptosporidium parvum oocysts in the presence of hydrated lime. Veterinary Record, 166, 297–30010.1136/vr.c1157Search in Google Scholar PubMed

Received: 2016-10-21
Revised: 2016-11-23
Accepted: 2016-11-24
Published Online: 2016-12-28
Published in Print: 2017-3-1

© W. Stefański Institute of Parasitology, PAS