Parasitology Research

, 105:413 | Cite as

Molecular epidemiology of human Blastocystis isolates in France

  • Laetitia Souppart
  • Giovanna Sanciu
  • Amandine Cian
  • Ivan Wawrzyniak
  • Frederic Delbac
  • Monique Capron
  • Eduardo Dei-Cas
  • Kenneth Boorom
  • Laurence Delhaes
  • Eric Viscogliosi
Original Paper

Abstract

Blastocystis sp. is the most common eukaryotic parasite in the intestinal tract of humans. Due to its strong impact in public health, in this study, we determined the frequency of different Blastocystis subtypes in patients in France. We hypothesized on the mode of transmission and tested a possible relationship between the subtype and symptomatic status. We obtained a total of 40 stool samples identified as positive for Blastocystis by microscopic examination of smears. Participants consisted of 25 symptomatic and 15 asymptomatic patients, for whom clinical and parasitological data were collected. For nested-polymerase chain reaction and genotyping, DNA was extracted directly from fecal samples or from fecal cultures. Morphological forms observed in fecal cultures were uncorrelated with symptomatic status. Genotyping using partial small subunit rRNA gene analysis identified a total of 43 Blastocystis isolates corresponding to 37 single infections and three mixed infections by two different subtypes. These 43 isolates belonged to five subtypes (1, 2, 3, 4, and 7) with predominance of subtype 3 (53.5%). Patient symptomatic status was uncorrelated with Blastocystis subtype.

References

  1. Abe N (2004) Molecular and phylogenetic analysis of Blastocystis isolates from various hosts. Vet Parasitol 120:235–242CrossRefPubMedGoogle Scholar
  2. Abe N, Nagoshi M, Takami K, Sawano Y, Yoshikawa H (2002) A survey of Blastocystis sp. in livestock, pets, and zoo animals in Japan. Vet Parasitol 106:203–212PubMedGoogle Scholar
  3. Abe N, Wu Z, Yoshikawa H (2003) Zoonotic genotypes of Blastocystis hominis detected in cattle and pigs by PCR with diagnostic primers and restriction fragment length polymorphism analysis of the small subunit ribosomal RNA gene. Parasitol Res 90:124–128PubMedGoogle Scholar
  4. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402CrossRefPubMedGoogle Scholar
  5. Arisue N, Hashimoto T, Yoshikawa H (2003) Sequence heterogeneity of the small subunit ribosomal RNA genes among Blastocystis isolates. Parasitology 126:1–9CrossRefPubMedGoogle Scholar
  6. Böhm-Gloning B, Knobloch J, Walderich B (1997) Five subgroups of Blastocystis hominis isolates from symptomatic and asymptomatic patients revealed by restriction site analysis of PCR-amplified 16 S-like rDNA. Trop Med Int Health 2:117–118CrossRefGoogle Scholar
  7. Boorom KF, Smith H, Nimri L, Viscogliosi E, Spanakos G, Parkar U, Li LH, Zhou XN, Ok UZ, Leelayoova S, Jones MS (2008) Oh my aching gut: irritable bowel syndrome, Blastocystis, and asymptomatic infection. BMC Parasit Vectors 1:40CrossRefGoogle Scholar
  8. Clark CG (1997) Extensive genetic diversity in Blastocystis hominis. Mol Biochem Parasitol 87:79–86CrossRefPubMedGoogle Scholar
  9. Dogruman-Al F, Dragci H, Yoshikawa H, Kurt Ö, Demirel M (2008) A possible link between subtype 2 and asymptomatic infections of Blastocystis hominis. Parasitol Res 103:685–689CrossRefPubMedGoogle Scholar
  10. Giacometti A, Cirioni O, Fiorentini A, Fortuna M, Scalise G (1999) Irritable bowel syndrome in patients with Blastocystis hominis infection. Eur J Clin Microbiol Infect Dis 18:436–439CrossRefPubMedGoogle Scholar
  11. Hoevers J, Holman P, Logan K, Hommel M, Ashford R, Snowden K (2000) Restriction-fragment-length polymorphism analysis of small subunit rRNA genes of Blastocystis hominis isolates from geographically diverse human hosts. Parasitol Res 86:57–61CrossRefPubMedGoogle Scholar
  12. Hunter PR, Hughes S, Woodhouse S, Syed Q, Verlander NQ, Chalmers RM, Morgan K, Nichols G, Beeching N, Osborn K (2004) Sporadic cryptosporidiosis case-control study with genotyping. Emerg Inf Dis 10:1241–1249Google Scholar
  13. Hussein EM, Hussein AM, Eida MM, Atwa MM (2008) Pathophysiological variability of different genotypes of human Blastocystis hominis Egyptian isolates in experimentally infected rats. Parasitol Res 102:853–860CrossRefPubMedGoogle Scholar
  14. Init I, Mak JW, Lokman HS, Yong HS (1999) Strain differences in Blastocystis isolates as detected by a single set of polymerase chain reaction primers. Parasitol Res 85:131–134CrossRefPubMedGoogle Scholar
  15. Jones WR (1946) The experimental infection of rats with Entamoeba hystolitica with a method for evaluating the anti-amoeboic properties of new compounds. Ann Trop Med Parasitol 40:130–140Google Scholar
  16. Jones MS, Whipps CM, Ganac RD, Hudson NR, Boroom K (2009) Association of Blastocystis subtype 3 and 1 with patients from an Oregon community presenting with chronic gastrointestinal illness. Parasitol Res 104:341–345CrossRefPubMedGoogle Scholar
  17. Kaneda Y, Horiki N, Cheng XJ, Fujita Y, Maruyama M, Tachibana H (2001) Ribodemes of Blastocystis hominis isolated in Japan. Am J Trop Med Hyg 65:393–396PubMedGoogle Scholar
  18. Katsarou-Katsari A, Vassalos CM, Tzanetou K, Spanakos G, Papadopoulou C, Vakalis N (2008) Acute urticaria associated with amoeboid forms of Blastocystis sp. subtype 3. Acta Derm Venereol 88:80–81CrossRefPubMedGoogle Scholar
  19. Leelayoova S, Siripattanapipong S, Thathaisong U, Naaglor T, Taamasri P, Piyaraj P, Mungthin M (2008) Drinking water: a possible source of Blastocystis spp. subtype 1 infection in schoolchildren of a rural community in central Thailand. Am J Trop Med Hyg 79:401–406PubMedGoogle Scholar
  20. Li LH, Zhang XP, Shan L, Yoshikawa H, Zhiliang W, Steinmann P, Utzinger J, Tong XM, Chen SH, Zhou XN (2007a) Cross-sectional surveys and subtype classification of human Blastocystis isolates from four epidemiological settings in China. Parasitol Res 102:83–90CrossRefPubMedGoogle Scholar
  21. Li LH, Zhou XN, Du ZW, Wang XZ, Wang LB, Jiang JY, Yoshikawa H, Steinmann P, Utzinger J, Wu Z, Chen JX, Chen SH, Zhang L (2007b) Molecular epidemiology of human Blastocystis in a village in Yunnan province, China. Parasitol Int 56:281–286CrossRefPubMedGoogle Scholar
  22. Menounos PG, Spanako G, Tegos N, Vassalos CM, Papadopoulou C, Vakalis C (2007) Direct detection of Blastocystis sp. in human faecal samples and subtype assignment using single strand conformational polymorphism and sequencing. Mol Cell Probes 22:24–29CrossRefPubMedGoogle Scholar
  23. Moe KT, Singh M, Howe J, Ho LC, Tan SW, Ng GC, Chen XQ, Yap EH (1996) Observations on the ultrastructure and viability of the cystic stage of Blastocystis hominis from human feces. Parasitol Res 82:439–444CrossRefPubMedGoogle Scholar
  24. Noël C, Peyronnet C, Gerbod D, Edgcomb VP, Delgado-Viscogliosi P, Sogin ML, Capron M, Viscogliosi E, Zenner L (2003) Phylogenetic analysis of Blastocystis isolates from different hosts based on the comparison of small-subunit rRNA gene sequences. Mol Biochem Parasitol 126:119–123CrossRefPubMedGoogle Scholar
  25. Noël C, Dufernez F, Gerbod D, Edgcomb VP, Delgado-Viscogliosi P, Ho LC, Singh M, Wintjens R, Sogin ML, Capron M, Pierce R, Zenner L, Viscogliosi E (2005) Molecular phylogenies of Blastocystis isolates from different hosts: implications for genetic diversity, identification of species, and zoonosis. J Clin Microbiol 43:348–355CrossRefPubMedGoogle Scholar
  26. Özyurt M, Kurt O, Molbak K, Nielsen HV, Haznedaroglu T, Stensvold CR (2008) Molecular epidemiology of Blastocystis infections in Turkey. Parasitol Int 57:300–306CrossRefPubMedGoogle Scholar
  27. Parkar U, Traub RJ, Kumar S, Mungthin M, Vitali S, Leelayoova S, Morris K, Thompson RCA (2007) Direct characterization of Blastocystis from faeces by PCR and evidence of zoonotic potential. Parasitology 134:359–367CrossRefPubMedGoogle Scholar
  28. Rivera WL (2008) Phylogenetic analysis of Blastocystis isolates from animal and human hosts in the Philippines. Vet Parasitol 156:178–182CrossRefPubMedGoogle Scholar
  29. Rivera WL, Tan MAV (2005) Molecular characterization of Blastocystis isolates in the Philippines by riboprinting. Parasitol Res 96:253–257CrossRefPubMedGoogle Scholar
  30. Salim HR, Kumar GS, Vellayan S, Mak JW, Anuar AK, Init I, Vennila GD, Saminathan R, Ramakrishnan K (1999) Blastocystis in animal handlers. Parasitol Res 85:1032–1033CrossRefGoogle Scholar
  31. Scicluna SM, Tawari B, Clark CG (2006) DNA barcoding of Blastocystis. Protist 157:77–85CrossRefPubMedGoogle Scholar
  32. Stark D, van Hal S, Marriott D, Elis J, Harkness J (2007) Irritable bowel syndrome: a review on the role of intestinal protozoa and the importance of their detection and diagnosis. Int J Parasitol 37:11–20CrossRefPubMedGoogle Scholar
  33. Stensvold CR, Brillowska-Dabrowska A, Nielsen HV, Arendrup MC (2006) Detection of Blastocystis hominis in unpreserved stool specimens by using polymerase chain reaction. J Parasitol 92:1081–1087CrossRefPubMedGoogle Scholar
  34. Stensvold CR, Arendrup MC, Jespersgaard C, Molbak K, Nielsen HV (2007a) Detecting Blastocystis using parasitologic and DNA-based methods: a comparative study. Diag Microbiol Infect Dis 59:303–307Google Scholar
  35. Stensvold CR, Suresh GK, Tan KSW, Thompson RCA, Traub RJ, Viscogliosi E, Yoshikawa H, Clark CG (2007b) Terminology for Blastocystis subtypes-a consensus. Trends Parasitol 23:93–96CrossRefPubMedGoogle Scholar
  36. Stensvold CR, Nielsen HV, Mølbak K, Smith HV (2009a) Pursuing the clinical significance of Blastocystis—diagnostic limitations. Trends Parasitol 25:23–29CrossRefPubMedGoogle Scholar
  37. Stensvold CR, Alfellani MA, Nørskov-Lauritsen S, Prip K, Victory EL, Maddox C, Nielsen HV, Clark CG (2009b) Subtype distribution of Blastocystis isolates from synanthropic and zoo animals and identification of a new subtype. Int J Parasitol 39:473–479CrossRefPubMedGoogle Scholar
  38. Stenzel DJ, Boreham PFL (1996) Blastocystis hominis revisited. Clin Microbiol Rev 9:563–584PubMedGoogle Scholar
  39. Suresh K, Smith HV, Tan TC (2005) Viable Blastocystis cysts in Scottish and Malaysian sewage samples. Appl Environ Microbiol 71:5619–5620CrossRefPubMedGoogle Scholar
  40. Tan KSW (2004) Blastocystis in humans and animals: new insights using modern methodologies. Vet Parasitol 126:121–144CrossRefPubMedGoogle Scholar
  41. Tan KSW (2008) New insights on classification, identification, and clinical relevance of Blastocystis spp. Clin Microbiol Rev 21:639–665CrossRefPubMedGoogle Scholar
  42. Tan TC, Suresh KG (2006) Predominance of amoeboid forms of Blastocystis hominis in isolates from symptomatic patients. Parasitol Res 98:189–193CrossRefPubMedGoogle Scholar
  43. Tan KSW, Singh M, Yap EH (2002) Recent advances in Blastocystis hominis research: hot spots in terra incognita. Int J Parasitol 32:789–804CrossRefPubMedGoogle Scholar
  44. Tan TC, Suresh KG, Smith HV (2008) Phenotypic and genotypic characterization of Blastocystis hominis isolates implicates subtype 3 as a subtype with pathogenic potential. Parasitol Res 104:85–93CrossRefPubMedGoogle Scholar
  45. Wong KHS, Ng GC, Lin RTP, Yoshikawa H, Taylor MB, Tan KSW (2008) Predominance of subtype 3 among Blastocystis isolates from a major hospital in Singapore. Parasitol Res 102:663–670CrossRefPubMedGoogle Scholar
  46. Xiao L, Fayer R, Ryan U, Upton SJ (2004) Cryptosporidium taxonomy: recent advances and implications for public health. Clin Microbiol Rev 17:72–97CrossRefPubMedGoogle Scholar
  47. Yakoob J, Jafri W, Jafir N, Khan R, Islam M, AsimBeg M, Zaman V (2004) Irritable bowel syndrome: in search of an etiology: role of Blastocystis hominis. Am J Trop Med Hyg 70:383–385PubMedGoogle Scholar
  48. Yan Y, Su S, Lai R, Liao H, Ye J, Li X, Luo X, Chen G (2006) Genetic variability of Blastocystis isolates in China. Parasitol Res 99:597–601CrossRefPubMedGoogle Scholar
  49. Yan Y, Su S, Ye J, Lai X, Lai R, Liao H, Chen G, Zhang R, Hou Z, Luo X (2007) Blastocystis sp. subtype 5: a possibly zoonotic genotype. Parasitol Res 101:1527–1532CrossRefPubMedGoogle Scholar
  50. Yoshikawa H, Nagono I, Yap EP, Singh M, Takahashi Y (1996) DNA polymorphism revealed by arbitrary primers polymerase chain reaction among Blastocystis strains isolated from humans, a chicken and a reptile. J Eukaryot Microbiol 43:127–30CrossRefPubMedGoogle Scholar
  51. Yoshikawa H, Nagano I, Wu Z, Yap EH, Singh M, Takahashi Y (1998) Genomic polymorphism among Blastocystis hominis strains and development of subtype-specific diagnostic primers. Mol Cell Probes 12:153–159CrossRefPubMedGoogle Scholar
  52. Yoshikawa H, Wu Z, Nagano I, Takahashi Y (2003) Molecular comparative studies among Blastocystis isolates obtained from humans and animals. J Parasitol 89:585–594CrossRefPubMedGoogle Scholar
  53. Yoshikawa H, Wu Z, Kimata I, Iseki M, Ali IKMD, Hossain MB, Zaman V, Haque R, Takahashi Y (2004) Polymerase chain reaction-based genotype classification among human Blastocystis hominis populations isolated from different countries. Parasitol Res 92:22–29CrossRefPubMedGoogle Scholar
  54. Yoshikawa H, Wu Z, Pandey K, Pandey BD, Sherchand JB, Yanagi T, Kanbara H (2009) Molecular characterization of Blastocystis isolates from children and rhesus monkeys in Kathmandu, Nepal. Vet Parasitol 16:295–300CrossRefGoogle Scholar
  55. Zhang X, Qiao JY, Zhou XJ, Yao FR, Wei ZC (2007) Morphology and reproductive mode of Blastocystis hominis in diarrhea and in vitro. Parasitol Res 101:43–51CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Laetitia Souppart
    • 1
  • Giovanna Sanciu
    • 1
  • Amandine Cian
    • 1
  • Ivan Wawrzyniak
    • 2
  • Frederic Delbac
    • 2
  • Monique Capron
    • 1
  • Eduardo Dei-Cas
    • 3
  • Kenneth Boorom
    • 4
  • Laurence Delhaes
    • 3
  • Eric Viscogliosi
    • 1
  1. 1.Inserm, U547, Institut Pasteur de LilleUniversité Lille Nord de FranceLille CedexFrance
  2. 2.Laboratoire Microorganismes: Génome et Environnement, UMR CNRS 6023Université Blaise PascalAubière CedexFrance
  3. 3.Institut Pasteur de Lille, Laboratoire d’Ecologie du Parasitisme (EA3609) and Faculté de Médecine, Centre Hospitalier Régional UniversitaireUniversité Lille Nord de FranceLilleFrance
  4. 4.Blastocystis Research FoundationCorvallisUSA

Personalised recommendations