Skip to main content
Springer Nature - PMC COVID-19 Collection logoLink to Springer Nature - PMC COVID-19 Collection
. 2016 Apr 1;61(2):406–411. doi: 10.1515/ap-2016-0054

First report of molecular identification of Cystoisospora suis in piglets with lethal diarrhea in Japan

Makoto Matsubayashi 227,327, Hideko Takayama 327, Masahiro Kusumoto 227, Misato Murata 327, Yuka Uchiyama 427, Masaya Kaji 527, Kazumi Sasai 127, Ryosaku Yamaguchi 227,627, Tomoyuki Shibahara 227,
PMCID: PMC7088846  PMID: 27078667

Abstract

Cystoisospora suis is a pathogen that causes diarrhea in pigs and can lead to serious disease. Species identification, especially by histopathological examination, is often difficult because of morphologically similar parasites such as Eimeria species. In this study, we used histopathological, bacteriological, virological, and parasitological methods to identify the cause of the disease in two piglets with severe diarrhea. Villous atrophy, diffuse necrosis, and flattening of mucosal epithelial cells were found in the ilea of examined piglets, and coccidian parasites were found in the cytoplasm of the epithelial cells. In some merozoites in the meronts, the presence of two nuclei indicated type 1 merozoites, characteristic of C. suis. According to Cystoisospora-specific PCR targeting the rRNA internal transcribed spacer 1 (ITS1) gene, the sequences of the products were 98.5% similar to those of C. suis. Escherichia coli (O149 serogroup) exhibiting a virulence factor profile (LT, STb, and EAST1 as toxins and F4 as a colonization factor) was detected in one piglet. No other bacteria or significant enteric viruses were found. Co-infection with C. suis and E. coli could imply aggravation of the disease, although further study is needed to assess the pathogenicity of this interaction. This study is the first to clarify by molecular analysis the sequences of C. suis detected in piglets in Japan.

Keywords: Cystoisospora suis, Escherichia coli, ITS1 gene, piglet

References

  1. Berberov EM, Zhou Y, Francis DH, Scott MA, Kachman SD, Moxley RA. Relative importance of heat-labile enterotoxin in the causation of severe diarrheal disease in the gnotobiotic piglet model by a strain of enterotoxigenic Escherichia coli that produces multiple enterotoxins. Infection and Immunity. 2004;72:3914–3924. doi: 10.1128/IAI.72.7.3914-3924.2004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Clark CG, Diamond LS. The Laredo strain and other ‘Entamoeba histolytica-like’ amoebae are Entamoeba moshkovskii. Molecular and Biochemical Parasitology. 1991;46:11–18. doi: 10.1016/0166-6851(91)90194-B. [DOI] [PubMed] [Google Scholar]
  3. Fairbrother JM, Nadeau E, Gyles CL. Escherichia coli in postweaning diarrhea in pigs: an update on bacterial types, pathogenesis, and prevention strategies. Animal Health Research Reviews. 2005;6:17–39. doi: 10.1079/AHR2005105. [DOI] [PubMed] [Google Scholar]
  4. Fairbrother JM, Ngeleka M. Extraintestinal Escherichia coli infections in pigs. In: Gyles C L, editor. Escherichia coli in domestic animals and humans. Wallingford, United Kingdom: CAB International; 1994. pp. 221–236. [Google Scholar]
  5. Joachim A, Ruttkowski B, Zimmermann M, Daugschies A, Mundt HC. Detection of Isospora suis (Biester and Murray 1934) in piglet faeces—comparison of microscopy and PCR. Journal of veterinary medicine. B. Infectious diseases and veterinary public health. 2004;51:140–142. doi: 10.1111/j.1439-0450.2004.00736.x. [DOI] [PubMed] [Google Scholar]
  6. Jones GW, Parker RJ, Parke CR. Coccidia associated with enteritis in grower pigs. Australian Veterinary Journal. 1985;62:319. doi: 10.1111/j.1751-0813.1985.tb14917.x. [DOI] [PubMed] [Google Scholar]
  7. Kim SY, Song DS, Park BK. Differential detection of transmissible gastroenteritis virus and porcine epidemic diarrhea virus by duplex RT-PCR. Journal of Veterinary Diagnostic Investigation. 2001;13:516–520. doi: 10.1177/104063870101300611. [DOI] [PubMed] [Google Scholar]
  8. Lindsay DS, Dubey JP, Santin-Duran M, Fayer R. Chapter 66: Coccidia and Other Protozoa. In: Zimmerman JJ, editor. Diseases of Swine. 10th ed. Iowa: Wiley-Blackwell; 2012. pp. 895–907. [Google Scholar]
  9. Lindsay DS, Neiger R, Hildreth M. Porcine enteritis associated with Eimeria spinosa Henry, 1931 infection. Journal of Parasitology. 2002;88:1262–1263. doi: 10.1645/0022-3395(2002)088[1262:PEAWES]2.0.CO;2. [DOI] [PubMed] [Google Scholar]
  10. Matsubayashi M, Carreno RA, Tani H, Yoshiuchi R, Kanai T, Kimata I, Uni S, Furuya M, Sasai K. Phylogenetic identification of Cystoisospora spp. from dogs, cats, and raccoon dogs in Japan. Veterinary Parasitology. 2011;176:270–274. doi: 10.1016/j.vetpar.2010.11.008. [DOI] [PubMed] [Google Scholar]
  11. Matsubayashi M, Suzuta F, Terayama Y, Shimojo K, Yui T, Haritani M, Shibahara T. Ultrastructural characteristics and molecular identification of Entamoeba suis isolated from pigs with hemorrhagic colitis: implications for pathogenicity. Parasitology Research. 2014;113:3023–3028. doi: 10.1007/s00436-014-3965-y. [DOI] [PubMed] [Google Scholar]
  12. Mengel H, Kruger M, Kruger MU, Westphal B, Swidsinski A, Schwarz S, Mundt HC, Dittmar K, Daugschies A. Necrotic enteritis due to simultaneous infection with Isospora suis and clostridia in newborn piglets and its prevention by early treatment with toltrazuril. Parasitology Research. 2012;110:1347–1355. doi: 10.1007/s00436-011-2633-8. [DOI] [PubMed] [Google Scholar]
  13. Mundt HC, Cohnen A, Daugschies A, Joachim A, Prosl H, Schmäschke R, Westphal B. Occurrence of Isospora suis in Germany, Switzerland and Austria. Journal of Veterinary Medicine B Infectious Diseases and Veterinary Public Health. 2005;52:93–97. doi: 10.1111/j.1439-0450.2005.00824.x. [DOI] [PubMed] [Google Scholar]
  14. Mundt H.-C., Joachim A., Daugschies A., Zimmermann M. Population biology studies on Isospora suis in piglets. Parasitology Research. 2003;90(0):S158–S159. doi: 10.1007/s00436-003-0926-2. [DOI] [PubMed] [Google Scholar]
  15. Mundt HC, Kruger M, Westphal B, Mengel H, Kruger MU, Dittmar K, Kuhnert Y, Daugschies A. Investigations into the synergic effect of postnatal experimental infection with Isospora suis and natural infection with Clostridium perfringens β 2 for the development of necrotic enteritis in piglets; 2008. p. 133. [Google Scholar]
  16. Rommel M. Parasitosen des Schweines, Protozoen. In: Eckert J, Kutzer E, Rommel M, Bürger H-J, Körting W, editors. Veterinärmedizinische Parasitologie. 4th ed. Berlin: Paul Parey; 1992. pp. 444–458. [Google Scholar]
  17. Ruttkowski B, Joachim A, Daugschies A. PCR-based differentiation of three porcine Eimeria species and Isospora suis. Veterinary Parasitology. 2001;95:17–23. doi: 10.1016/S0304-4017(00)00408-8. [DOI] [PubMed] [Google Scholar]
  18. Saitoh Y, Hattori J. Survey of porcine neonatal coccidium Isospora suis (Biester and Murray 1934) in Japan. Proceedings of Japan Pig Veterinary Society. 2007;50:7–8. [Google Scholar]
  19. Samarasinghe B, Johnson J, Ryan U. Phylogenetic analysis of Cystoisospora species at the rRNA ITS1 locus and development of a PCR-RFLP assay. Experimental Parasitology. 2008;118:592–595. doi: 10.1016/j.exppara.2007.10.015. [DOI] [PubMed] [Google Scholar]
  20. Stuart BP, Gosser HS, Allen CB, Bedell DM. Coccidiosis in swine: dose and age response to Isospora suis. Canadian Journal of Comparative Medicine. 1982;46:317–320. [PMC free article] [PubMed] [Google Scholar]
  21. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution. 2011;28:2731–2739. doi: 10.1093/molbev/msr121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Vu-Khac H, Holoda E, Pilipcinec E, Blanco M, Blanco JE, Dahbi G, Mora A, López C, González EA, Blanco J. Serotypes, virulence genes, intimin types and PFGE profiles of Escherichia coli isolated from piglets with diarrhoea in Slovakia. Veterinary Journal. 2007;174:176–187. doi: 10.1016/j.tvjl.2006.05.019. [DOI] [PubMed] [Google Scholar]

Articles from Acta Parasitologica are provided here courtesy of Nature Publishing Group

RESOURCES