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Analysis of the ITS region and partial ssu and lsu rRNA genes of Blastocystis and Proteromonas lacertae

Published online by Cambridge University Press:  25 April 2005

J. D. HOEVERS  and
K. F. SNOWDEN
J. D. HOEVERS
Affiliation:
Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, TX 77843-4467, USA
K. F. SNOWDEN
Affiliation:
Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, TX 77843-4467, USA
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Abstract

Blastocystis is a common single-celled enteric parasite found in a large variety of hosts. Recent molecular analysis supports the concept that this eukaryotic organism is a stramenopile most closely related to Proteromonas lacertae, a parasite of reptiles. In this study, the internal transcribed spacer region, partial small subunit rRNA and large subunit rRNA genes from 7 Blastocystis isolates (5 human, 1 pig and 1 sheep), and a Proteromonas lacertae isolate were amplified by PCR, cloned and sequenced. Blastocystis was found to be a typical eukaryote with both ITS1 and ITS2 regions present. Phylogenetic analysis based on the entire PCR amplicon revealed that the Blastocystis isolates did not segregate according to host or geographic origin. The highest sequence identities with the conserved Blastocystis 5·8S rDNA sequence were with the stramenopiles Fibrocapsa japonica, Chattonella marina, Cylindrotheca closterium and Hyphochytrium catenoides. The most parsimonious tree based on the 5·8S rDNA sequence from P. lacertae, 11 other stramenopiles, 2 fungi, 3 algae and 3 alveolates showed Blastocystis positioned within the stramenopiles, with P. lacertae as its closest relative. This work therefore supports the hypothesis that Blastocystis is most closely related to P. lacertae, and that it should be regarded as an unusual stramenopile.

Keywords

BlastocystisProteromonas lacertaeITS5·8Sssu rRNAlsu rRNAstramenopilemolecular phylogeny
Type
Research Article
Information
Parasitology , Volume 131 , Issue 2 , August 2005 , pp. 187 - 196
Copyright
2005 Cambridge University Press

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References

REFERENCES

ABE, N. ( 2004). Molecular and phylogenetic analysis of Blastocystis isolates from various hosts. Veterinary Parasitology 120, 235–242.CrossRefGoogle Scholar
ABE, N., NAGOSHI, M., TAKAMI, K., SAWANO, Y. & YOSHIKAWA, H. ( 2002). A survey of Blastocystis sp. in livestock, pets, and zoo animals in Japan. Veterinary Parasitology 106, 203–212.Google Scholar
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. Parasitology Research 90, 124–128.Google Scholar
ARISUE, N., HASHIMOTO, T. & YOSHIKAWA, H. ( 2003). Sequence heterogeneity of the small subunit ribosomal RNA genes among Blastocystis isolates. Parasitology 126, 1–9.CrossRefGoogle Scholar
ARISUE, N., HASHIMOTO, T., YOSHIKAWA, H., NAKAMURA, Y., NAKAMURA, G., NAKAMURA, F., YANO, T. & HASEGAWA, M. ( 2002). Phylogenetic position of Blastocystis hominis and of stramenopiles inferred from multiple molecular sequence data. Journal of Eukaryotic Microbiology 49, 42–53.CrossRefGoogle Scholar
ALLARD, M. W. & HONEYCUTT, R. L. ( 1991). Ribosomal DNA variation within and between species of rodents, with emphasis on the genus Onychomys. Molecular and Biochemical Evolution 8, 71–84.Google Scholar
ALTSCHUL, S. F., GISH, W., MILLER, W., MYERS, E. W. & LIPMAN, D. J. ( 1990). Basic local alignment search tool. Journal of Molecular Biology 215, 403–410.CrossRefGoogle Scholar
ASHFORD, R. W. & ATKINSON, E. A. ( 1992). Epidemiology of Blastocystis hominis infection in Papua New Guinea: age-prevalence and associations with other parasites. Annals of Tropical Medicine and Parasitology 86, 129–136.CrossRefGoogle Scholar
BALDWIN, B. G. ( 1992). Phylogenetic utility of the internal transcribed spacers of ribosomal DNA in plants. Molecular and Phylogenetic Evolution 1, 3–16.CrossRefGoogle Scholar
BARNARD, S. M. & UPTON, S. J. ( 1994). A Veterinary Guide to the Parasites of Reptiles: Protozoa, 1st Edn. Krieger Publishing Company, Melbourne, Florida.
BOREHAM, P. F. L. & STENZEL, D. J. ( 1998). Blastocystis. In Zoonoses: Biology, Clinical Practice, and Public Health Control (ed. Palmer, S. R., Soulsby, L. & Simpson, D. I. H.), pp. 625–632. Oxford University Press, Oxford.
CAVALIER-SMITH, T. ( 1998). A revised six-kingdom system of life. Biological Reviews of the Cambridge Philosophical Society 73, 203–266.CrossRefGoogle Scholar
COLLINS, N. E. & ALLSOPP, B. A. ( 1999). Theileria parva ribosomal internal transcribed spacer sequences exhibit extensive polymorphism and mosaic evolution: application to the characterization of parasites from cattle and buffalo. Parasitology 118, 541–551.CrossRefGoogle Scholar
FAITH, D. P. ( 1991). Cladistic permutation tests for monophyly and non-monophyly. Cladistics 40, 366–375.Google Scholar
FELSENSTEIN, J. ( 1985). Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39, 783–791.CrossRefGoogle Scholar
HO, L. C., ARMIUGAM, A., JEYASEELAN, K., YAP, E. H. & SINGH, M. ( 2000). Blastocystis elongation factor-1 alpha: genomic organization, taxonomy and phylogenetic relationships. Parasitology 121, 135–144.CrossRefGoogle Scholar
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. Parasitology Research 86, 57–61.CrossRefGoogle Scholar
JOHNSON, A. M., THANOU, A., BOREHAM, P. F. L. & BAVERSTOCK, P. R. ( 1989). Blastocystis hominis: phylogenetic affinities determined by rRNA sequence comparison. Experimental Parasitology 68, 283–288.CrossRefGoogle Scholar
KATIYAR, S. K., VISVESVARA, G. S. & EDLIND, T. D. ( 1995). Comparisons of ribosomal RNA sequences from amitochondrial protozoa: implications for processing, mRNA binding and paromomycin susceptibility. Gene 152, 27–33.CrossRefGoogle Scholar
LEIPE, D. D., TONG, S. M., GOGGIN, C. L., SLEMENDA, S. B., PIENIAZEK, N. J. & SOGIN, M. L. ( 1996). 16S-like rDNA sequences from Developayella elegans, Labyrinthuloides haliotidis, and Proteromonas lacertae confirm that the stramenopiles are a primarily heterotrophic group. European Journal of Protistology 32, 449–459.CrossRefGoogle Scholar
MCLAUGHLIN, G. L., HOWE, D. K., BIGGS, D. R., SMITH, A. R., LUDWINSKI, P., FOX, B. C., TRIPATHY, N., FRASCH, C. E., WENGER, J. D., CAREY, R. B., HASSEN-KING, M. & VODKIN, H. ( 1993). Amplification of rDNA loci to detect and type Neisseria meningitidis and other eubacteria. Molecular and Cellular Probes 7, 7–17.CrossRefGoogle Scholar
MOE, K., SINGH, T. M., HOWE, J., HO, L. C., TAN, S. W., CHEN, X. Q., NG, G. C. & YAP, E. H. ( 1997). Experimental Blastocystis hominis infection in laboratory mice. Parasitology Research 83, 319–325.CrossRefGoogle Scholar
NAKAMURA, Y., HASHIMOTO, T., YOSHIKAWA, H., KAMAISHI, T., NAKAMURA, F., OKAMOTO, K. & HASEGAWA, M. ( 1996). Phylogenic position of Blastocystis hominis that contains cytochrome-free mitochondria, inferred from the protein phylogeny of elongation factor –?1. Molecular and Biochemical Parasitology 77, 241–245.CrossRefGoogle Scholar
NICKRENT, D. & SARGENT, M. ( 1991). An overview of the secondary structure of the V4 region of the eukaryotic small subunit ribosomal RNA. Nucleic Acids Research 19, 227–235.CrossRefGoogle Scholar
NOËL, C., DUFERNEZ, F., GERBOD, D., EDGCOMB, V. P., DELGADO-VISCOGLIOSI, P., HO, L. C., SINGH, M., WINTJENS, R., SOGIN, M. L., CAPRON, M., PIERCE, R., ZENNER, L. & VISCOGOGLIOSI, E. ( 2005). Molecular phylogenies of Blastocystis isolates from different hosts: implications for genetic diversity, identification of species, and zoonosis. Journal of Clinical Microbiology 43, 348–355.CrossRefGoogle Scholar
NOËL, C., PEYRONNET, C., GERBOD, D., EDGCOMB, V., DELGADO-VISCOGLIOSI, P., SOGIN, M., 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. Molecular and Biochemical Parasitology 126, 119–123.CrossRefGoogle Scholar
OLIVIER, C., VAN DE PAS, C., LEPP, P., YODER, K. & RELMAN, D. A. ( 2001). Sequence variability in the first internal transcribed spacer region within and among Cyclospora species is consistent with polymorphism. International Journal for Parasitology 31, 1475–1487.CrossRefGoogle Scholar
PAKANDL, M. ( 1992). An experimental transmission of porcine strains of Blastocystis sp. in mice and gerbils. Folia Parasitology 39, 383–386.Google Scholar
PATTERSON, D. J. ( 1989). Stramenopiles: chromophytes from a protistan perspective. In The Chromophyte Algae: Problems and Perspectives (ed. Green, J. C., Leadbeater, B. S. C. & Diver, W. L.), pp. 357–379. Clarendon Press, Oxford.
PEARSON, W. R. ( 1997). Effective protein sequence comparison. Methods in Enzymology 266, 227–258.Google Scholar
PÉLANDAKIS, M., SERRE, S. & PERNIN, P. ( 2000). Analysis of the 5·8S rRNA gene and the internal transcribed spacers in Naegleria spp. and in N. fowleri. Journal of Eukaryotic Microbiology 47, 116–121.CrossRefGoogle Scholar
POSADA, D. & CRANDALL, K. L. ( 1998). Model test: testing the model of DNA substitution. Bioinformatics 14, 817–818.CrossRefGoogle Scholar
SILBERMAN, J. D., SOGIN, M. L., LEIPE, D. D. & CLARK, C. G. ( 1996). Human parasite finds taxonomic home. Nature, London 380, 398.CrossRefGoogle Scholar
SNOWDEN, K., LOGAN, K., BLOZINSKI, C., HOEVERS, J. & HOLMAN, P. ( 2000). Restriction fragment length polymorphism analysis of small subunit rRNA genes of Blastocystis isolates from animal hosts. Parasitology Research 86, 62–66.CrossRefGoogle Scholar
SOM, I., AZAM, A. & BHATTACHARYA, A. ( 2000). Inter and intra-strain variation in the 5·8S ribosomal RNA and ITS sequences of Entamoeba histolytica and comparison with Entamoeba dispar, Entamoeba moshkovskii and Entamoeba invadens. International Journal for Parasitology 30, 723–728.CrossRefGoogle Scholar
STENZEL, D. J. & BOREHAM, P. F. L. ( 1996). Blastocystis hominis revisited. Clinical Microbiology Reviews 9, 563–584.Google Scholar
SWOFFORD, D. L. ( 1996). PAUP: phylogenetic analysis using parsimony, version 4.0. Sinauer Associates, Sunderland, Massachusetts.
TAN, K., SINGH, M. & YAP, E. ( 2002). Recent advances in Blastocystis hominis research: hot spots in terra incognita. International Journal for Parasitology 32, 789–804.CrossRefGoogle Scholar
THATHAISONG, U., WORAPONG, J., MUNGTHIN, M., TAN-ARIYA, P., VIPUTTIGUL, K., SUDATIS, A., NOONAI, A. & LEELAYOOVA, S. ( 2003). Blastocystis isolates from a pig and a horse are closely related to Blastocystis hominis. Journal of Clinical Microbiology 41, 967–975.CrossRefGoogle Scholar
VAN DE PEER, Y., DE RIJK, P., WUYTS, J., WINKELMANS, T. & DE WACHTER, R. ( 2000). The European Small Subunit Ribosomal RNA database. Nucleic Acids Research 28, 175–176.CrossRefGoogle Scholar
VAN DER AUWERA, G. & DE WACHTER, R. ( 1998). Structure of the large subunit rDNA from a diatom, and comparison between small and large subunit ribosomal RNA for studying stramenopile evolution. Journal of Eukaryotic Microbiology 6, 521–527.CrossRefGoogle Scholar
WHITE, T. J., BRUNS, T., LEE, S. & TAYLOR, J. ( 1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In PCR Protocols: a Guide to Methods and Applications (ed. Innis, M. A., Gelfand, D. H., Sninsky, J. J. & White, T. J.), pp. 315–322. Academic Press, San Diego, California.CrossRef
YOSHIKAWA, H., MORIMOTO, K., WU, Z., SINGH, M. & HASHIMOTO, T. ( 2004). Problems in speciation in the genus Blastocystis. Trends in Parasitology 20, 251–255.CrossRefGoogle Scholar
YOSHIKAWA, H., NAGANO, I., WU, Z., YAP, E. H., SINGH, M. & TAKAHASHI, Y. ( 1998). Genomic polymorphism among Blastocystis hominis strains and development of subtype-specific diagnostic primers. Molecular and Cellular Probes 12, 153–159.CrossRefGoogle Scholar
ZIERDT, C. H. ( 1991). Blastocystis hominis – past and future. Clinical Microbiology Reviews 4, 61–79.CrossRefGoogle Scholar