Parasitology Research

, 105:949 | Cite as

High prevalence of subtype 4 among isolates of Blastocystis hominis from symptomatic patients of a health district of Valencia (Spain)

  • M. Victoria Domínguez-Márquez
  • Remedios Guna
  • Carlos Muñoz
  • M. Teresa Gómez-Muñoz
  • Rafael BorrásEmail author
Original Paper


In order to know the genetic diversity of Blastocystis hominis from a health district of Valencia (Spain) 51 clinical isolates from symptomatic patients,31 axenic and 20 monoxenic, were ribotyped by analysing the restriction fragment length polymorphism (RFLP) of amplicons obtained by polymerase chain reaction (PCR) of small-subunit of ribosomal DNA genes (SSU-rDNA). For this purpose, DNA was subjected to two independent PCR (RD3-RD5, F1-R1) and to three independent treatments with restrictases (AluI, HinfI and RsaI). The digested DNA was separated electrophoretically, the isolates were clustered into ribotypes (ribodemes, RD3-RD5; subgroups, F1-R1) according to their profiles and the results were translated into genetic subtypes (ST) proposed by a consensus terminology. The results show that the isolates studied are an heterogeneous population and that both PCR-RFLP SSU-rDNA protocols have a similar discriminative power, since it allowed the ribotyping of all isolates and their clustering into four demes: ribodemes 1, 3 and 3-r and 6, which include isolates belonging to subgroup III, IV, V and V-r, respectively; which were assigned to ST1 (2%), ST2 (3.9%) and ST4 (94.1%). The most common of which is a zoonotic subtype (Blastocystis ratti) which includes, according to recent studies, non-pathogenic and pathogenic variants.


Restriction Fragment Length Polymorphism Health District Restriction Fragment Length Polymorphism Pattern Restriction Fragment Length Polymorphism Profile Independent Polymerase Chain Reaction 
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  1. Abe N, Nagoshi M, Takami K, Sawamo Y, Yoshikawa H (2002) A survey of Blastocystis sp. in livestock, pets, and zoo animals in Japan. Vet Parasitol 106:203–212PubMedGoogle Scholar
  2. Abe N, Wu Z, Yoshikawa H (2003) Molecular characterization of Blastocystis isolates from primates. Vet Parasitol 113:221–225CrossRefGoogle Scholar
  3. Belova LM, Kostenko LA (1990) Blastocystis galli sp. n. (Protista: Rhizopodea) from the intestine of domestic hens. Parazitologiia 24:164–168 in RussianPubMedGoogle Scholar
  4. 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 2:771–778PubMedCrossRefGoogle Scholar
  5. Boreham PFL, Stenzel DJ (1993) Blastocystosis in humans and animals: morphology, biology, and epizootiology. Adv Parasitol 32:1–70PubMedCrossRefGoogle Scholar
  6. Carbajal JA, Villar J, Lanuza MD, Esteban JG, Muñoz C, Borrás R (1997a) Significación clínica de la parasitación por Blastocystis hominis: estudio epidemiólogico. Med Clín (Bar) 108:606–612Google Scholar
  7. Carbajal JA, Del Castillo L, Lanuza MD, Villar J, Borrás R (1997b) Karyotypic diversity among Blastocystis hominis isolates. Int J Parasitol 27:941–945PubMedCrossRefGoogle Scholar
  8. Cassano N, Scoppio BM, Loviglio MC, Vena GA (2005) Remission of delayed pressure urticaria after erradication of Blastocystis hominis. Acta Derm Venereol 85:357–358PubMedGoogle Scholar
  9. Chen XQ, Singh M, Ho LC, Tan SW, Ng GC, Moe KT, Yap EH (1997) Description of a Blastocystis specie from Rattus norvegicus. Parasitol Res 83:79–83CrossRefGoogle Scholar
  10. Clark CG (1997) Extensive genetic diversity in Blastocysits hominis. Mol Bioch Parasitol 87:79–83CrossRefGoogle Scholar
  11. Clark CG (2000) Cryptic genetic variation in parasitic protozoa. J Med Microbiol 49:489–491PubMedGoogle Scholar
  12. Duda A, Stenzel DJ, Boreham PFL (1998) Detection of Blastocystis sp. in domestic dogs and cats. Vet Parasitol 76:9–17PubMedCrossRefGoogle Scholar
  13. 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 86:57–61PubMedCrossRefGoogle Scholar
  14. 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 102:853–860PubMedCrossRefGoogle Scholar
  15. Iguchi A, Ebisu A, Nagata S, Saitou Y, Yoshikawa H, Iwatani S, Kimata I (2007) Infectivity of different genotypes of human Blastocystis hominis isolates in chickens and rats. Parasitol Int 56:107–112PubMedCrossRefGoogle Scholar
  16. Iguchi A, Yoshikawa H, Yamada M, Kimata I, Arizono N (2009) Expression of interferon gamma and proinflammatory cytokines in the cecal mucosa or rats experimentally infected with Blastocystis sp. strain RN94-9. Parasitol Res . doi: 10.007/s00436-009-1373-5 PubMedGoogle 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. Krylov MV, Belova LM (1997) Blastocystis from primates. Parazitologiia 31:341–345 in RussianPubMedGoogle Scholar
  19. Lakhanpal S, Cohen SB, Fleischmann RM (1991) Reactive arthritis from Blastocystis hominis. Arthritis Rheum 34:251–253PubMedCrossRefGoogle Scholar
  20. Lanuza MD, Carbajal JA, Borrás R (1996) Identification of surface coat carbohydrates in Blastocystis hominis by lectin probes. Int J Parasitol 26:527–532PubMedCrossRefGoogle Scholar
  21. Lanuza MD, Carbajal JA, Villar J, Borrás R (1997) Description of an improved method for Blastocystis hominis culture and axenization. Parasitol Res 83:60–63PubMedCrossRefGoogle Scholar
  22. Lanuza MD, Carbajal JA, Villar J, Mir MD, Borrás R (1999) Soluble-protein and antigenic heterogeneity in axenic Blastocystis hominis isolates: pathogenic implications. Parasitol Res 85:93–97PubMedCrossRefGoogle Scholar
  23. Long HY, Handschack A, König W, Ambrosch A (2001) Blastocystis hominis modulates immune responses and cytokine release in colonic epithelial cells. Parasitol Res 87:1029–1030Google Scholar
  24. Mansour NS, Mikhail EN, El Masry NA, Sabry AG, Mohareb EW (1995) Biochemical characterization of human isolates of Blastocystis hominis. J Med Microbiol 42:304–307PubMedCrossRefGoogle Scholar
  25. Navarro C, Domínguez-Márquez MV, Garijo-Toledo M, Vega-García S, Fernández- Barredo S, Pérez-Gracia T, García A, Borrás R, Gómez-Muñoz MT (2008) High prevalence of Blastocystis sp. in pigs reared under intensive growing systems: frequency of ribotypes and associated risk factors. Vet Parasitol 153:347–358PubMedCrossRefGoogle Scholar
  26. Parkar U, Traub RJ, Kumar S, Mungthin M, Vitali S, Leelayoova S, Morris K, Thomson RCA (2007) Direct characterization of Blastocystis from faeces by PCR and evidence of zoonotic potential. Parasitology 134:359–367PubMedCrossRefGoogle Scholar
  27. Quílez J, Clavel A, Sánchez-Acedo C, Causapé AC, Albalá F (1993) Detection of Blastocystis sp. in pigs in Aragon (Spain). Vet Parasitol 56:345–348CrossRefGoogle Scholar
  28. Singh M, Ho LC, Yap AL, GC NG, Tan SW, Moe KT, Yap EH (1996) Axenic culture of reptilian Blastocystis isolates in monophasic medium and speciation by karyotypic typing. Parasitol Res 82:165–169PubMedCrossRefGoogle Scholar
  29. Stensvold CR, Suresh GK, Tan KS, Thompson RC, Traub RJ, Viscogliosi E, Yoshikawa H, Clark CG (2007) Terminology for Blastocystis subtypes—a consensus. Trends Parasitol 23:93–96PubMedCrossRefGoogle Scholar
  30. Stenzel DJ, Boreham PFL (1996) Blastocystis hominis revisited. Clin Microbiol Rev 9:563–584PubMedGoogle Scholar
  31. Taamasri P, Mungthin M, Rangsin R, Tongupprakarn B, Areekul W, Leelayoova S (2000) Transmission of intestinal blastocystosis related to the equality of drinking water. Southeast Asian J Trop Med Public Health 31:112–117PubMedGoogle Scholar
  32. Tan KSW (2008) New insights on classification, identification, and clinical relevance of Blastocystis spp. Clinical Microbiol Rev 21:639–665CrossRefGoogle Scholar
  33. Tan TC, Suresh KG, Smith HV (2008) Phenotypic and genotypic characterization of Blastocystis hominis isolates implicated subtype 3 as a subtype with pathogenic potential. Parasitol Res 104:85–93PubMedCrossRefGoogle Scholar
  34. Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DH, Swaminathan B (1995) Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol 33:2223–2239Google Scholar
  35. Teow WL, Zaman V, Ng GC, Chan YC, Yap EH, Howe J, Gopalakrishnakone P, Singh M (1991) Blastocystis species from the sea-snake, Lapemis hardwickii (Serpentes: Hydrophiidae). Int J Parasitol 6:723–726CrossRefGoogle Scholar
  36. Thathaisong U, Warapong J, Mungthin M, Tan-Ariya P, Viputtigul K, Sudatis A, Noonai A, Leelayoova S (2003) Blastocystis isolates from pig and horse are closely related to Blastocystis hominis. J Clin Microbiol 41:967–975PubMedCrossRefGoogle Scholar
  37. Windsor JJ, MacFarlane L, Hughes-Tapa G, Jones SKA, Whiteside TM (2002) Incidence of Blastocystis hominis in faecal samples submitted for routine microbiological analysis. Br J Biomed Sci 59:154–157PubMedGoogle Scholar
  38. Wong KHS, Ng GC, Lin RTP, Yoshikawa H, Taylor MB, Tan KSW (2007) Predominance of subtype 3 among Blastocystis isolates from a major hospital in Singapore. Parasitol Res 102:663–670PubMedCrossRefGoogle Scholar
  39. Yan Y, Su S, Lai R, Liao H, Ye J, Li X, Luo X, Chen G (2006) Genetic variability of Blastocystis hominis isolates in China. Parasitol Res 99:597–601PubMedCrossRefGoogle Scholar
  40. 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 12:153–159PubMedCrossRefGoogle Scholar
  41. Yoshikawa H, Abe N, Iwasawa M, Kitano S, Nagano I, Wu Z, Takahashi Y (2000) Genomic analysis of Blastocystis hominis strains isolated from two long-term health care facilities. J Clin Microbiol 38:1324–1330PubMedGoogle Scholar
  42. Yoshikawa H, Wu Z, Nagano I, Takahashi Y (2003) Molecular comparative studies among Blastocystis isolates obtained from humans and animals. J Parasitol 89:585–594PubMedCrossRefGoogle Scholar
  43. Yoshikawa H, Abe N, Wu Z (2004a) PCR-based identification of zoonotic isolates of Blastocystis from mammals and birds. Microbiology 150:1147–1151PubMedCrossRefGoogle Scholar
  44. Yoshikawa H, Wu Z, Kimata I, Iseki M, Karim MDAI, Hossain MB, Zaman V, Haque R, Takahashi Y (2004b) Polymerase chain reaction-based genotype classification among human Blastocystis hominis populations isolated from different countries. Parasitol Res 92:22–29PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • M. Victoria Domínguez-Márquez
    • 1
    • 3
  • Remedios Guna
    • 2
    • 3
  • Carlos Muñoz
    • 3
  • M. Teresa Gómez-Muñoz
    • 4
  • Rafael Borrás
    • 3
    • 5
    Email author
  1. 1.Servicio de MicrobiologíaHospital Universitario de La RiberaAlziraSpain
  2. 2.Unidad de MicrobiologíaConsorcio Hospital General UniversitarioValenciaSpain
  3. 3.Departamento de Microbiología y Ecología, Facultad de Medicina y OdontologíaUniversidad de ValenciaValenciaSpain
  4. 4.Departamento de Sanidad AnimalUniversidad Cardenal Herrera-CEUMoncadaSpain
  5. 5.Servicio de MicrobiologíaHospital Clínico UniversitarioValenciaSpain

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