Abstract
Extensive genomic polymorphism has been demonstrated among morphologically identical Blastocystis isolates. To this end, 32 Blastocystis isolates from the Philippines (12 from humans, 12 from pigs and 8 from chickens) were analyzed genetically by riboprinting or restriction fragment length polymorphism (RFLP) analysis of polymerase chain reaction (PCR) amplified small subunit rDNA. Three distinct riboprint patterns were observed from the HinfI digestion, while four patterns resulted from the RsaI digestion of Blastocystis SSU rDNA. Restriction fragment profiles between Blastocystis isolates from different hosts were generally different from each other. However, Blastocystis isolates within each host group were practically the same. Cluster analysis of the riboprint patterns revealed seven distinct groups of the Blastocystis isolates, including a zoonotic strain. These results demonstrate the genetic heterogeneity of Blastocystis in the Philippines and a support to the idea of the organism’s zoonotic potential.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abe N, Wu Z, Yoshikawa H (2003a) Molecular characterization of Blastocystis isolates from birds by PCR with diagnostic primers and restriction fragment length polymorphism analysis of the small subunit RNA gene. Parasitol Res 89: 393–396
Abe N, Wu Z, Yoshikawa H (2003b) Molecular characterization of Blastocystis isolates from primates. Vet Parasitol 113: 321–325
Abe N, Wu Z, Yoshikawa H (2003c) Zoonotic genotypes of Blastocystis hominis detected in cattle and pigs by PCR with diagnostic primers and restriction fragment length polymorsphism analysis of the small subunit ribosomal RNA gene. Parasitol Res 90: 124–128
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 16S-like rDNA. Trop Med Int Health 2: 771–778
Boreham PFL, Stenzel DJ (1993) Blastocystis in humans and animals: morphology, biology, and epizootiology. Adv Parasitol 32: 1–70
Clark CG (1997a) Extensive genetic diversity in Blastocystis hominis hominis. Mol Biochem Parasitol 87: 79–83
Clark CG (1997b) Riboprinting: a tool for the study of genetic diversity in microorganisms. J Euk Microbiol 44: 277–283
Diamond LS (1983) Lumen dwelling protozoa: Entamoeba, Trichomonads, and Giardia. In: Jensen JB (ed) In Vitro Cultivation of Protozoan Parasites. CRC Press, Boca Raton FL, pp 65–109
Kovach W (2002) Multivariate Statistical Package version 3.1d for Windows software. Kovach Computing Services, Wales
Rivera WL, Tachibana H, Silva-Tahat MRA, Uemura H, Kanbara H (1996) Differentiation of Entamoeba histolytica and E. dispar DNA from cysts present in stool specimens by polymerase chain reaction: its field application in the Philippines. Parasitol Res 82: 585–589
Rivera WL, Tachibana H, Kanbara H (1998) Field study on the distribution of Entamoeba histolytica and E. dispar in the Northern Philippines as detected by the polymerase chain reaction. Am J Trop Med Hyg 59: 916–921
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: 1032–1033
Stenzel DJ, Boreham PFL (1996) Blastocystis hominis revisited. Clin Microbiol Rev 9: 563–584
Tan KSW, Singh M, Yap EH (2002) Recent advances in Blastocystis hominis research: hot spots in terra incognita. Int J Parasitol 32: 789–804
Thathaisong U, Worapong J, Mungthin M, Tan-Ariya P, Viputtigul K, Sudatis A, Noonai A, Leelayoova S (2003) Blastocystis isolates from a pig and horse are closely related to Blastocystis hominis. J Clin Microbiol 41: 967–975
Yoshikawa H, Nagano I, Yap EH, 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 Euk Microbiol 43: 127–130
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: 153–159
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: 1324–1330
Yoshikawa H, Wu Z, Nagano I, Takahashi Y (2003) Molecular comparative studies among Blastocystis isolates obtained from humans and animals. J Parasitol 89: 585–594
Yoshikawa H, Abe N, Wu Z (2004a) PCR-based identification of zoonotic isolates of Blastocystis from mammals and birds. Microbiology 150: 1147–1151
Yoshikawa H, Wu Z, Kimata I, Iseki M, Ali IKMD, 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: 22–29
Acknowledgements
We thank Dr. Hisao Yoshikawa for the gift of Blastocystis genomic DNAs from reference cultures. This work was supported by a grant from the Natural Sciences Research Institute of the University of the Philippines (BIO-03-2-02) to W.L.R.
Author information
Authors and Affiliations
Corresponding author
Additional information
Declaration: The experiments conducted in this study comply with the current laws of the Philippines.
Rights and permissions
About this article
Cite this article
Rivera, W.L., Tan, M.A.V. Molecular characterization of Blastocystis isolates in the Philippines by riboprinting. Parasitol Res 96, 253–257 (2005). https://doi.org/10.1007/s00436-005-1344-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-005-1344-4