Skip to main content

Advertisement

Log in

Preliminary genomic characterization of microsporidian Nosema bombycis

  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

In order to characterize the genome of Nosema bombycis, the techniques of karyotyping, pulsed field gel electrophoresis, and polymerase chain reaction were applied. Nosema genomic DNA moved as 23 kb fragment on a standard agarose gel. The karyotype showed four chromosomes, the molecular karyotyping by pulsed field gel electrophoresis also showed four chromosomes. Arbitrarily primed polymerase chain reaction (PCR) with various primers showed amplification products of sizes ranging from 1.6 to 0.15 kb. Polymerase chain reaction with specific primer showed an amplification product of approximately 315 nucleotides. The DNA hybridizations are discussed. This is the first report of its kind on microsporidian Nosema bombycis. The current data can play a major role in elucidating the molecular biology of this parasite.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Adam, R.D., Theodore, E.N. & Thomas, E. Wellems. 1998 The Giardia lamblia trophozcite contains 3 sets of closely related chromosomes. Nucleic Acids Research 16, 4555-4567.

    Google Scholar 

  • Baig, M., Datta, R.K., Nataraju, B., Samson, M.V. & Sivaprasad, V. 1992 Protein A linked Latex antisera test for the detection of Nosema bomycis Naegeli spores. Journal of Invertebrate Pathology 60, 312-313.

    Article  Google Scholar 

  • Baker, M.D., Vossbrinck, C.R. & Undeen, A.H. 1994 Phylogenetic relationships among Vairimorpha and Nosema species (Microspora) based on ribosomal RNA sequence data. Journal of Invertebrate Pathology 64, 100-106.

    Article  CAS  Google Scholar 

  • Biderre, C., Pages, M., Metenier, G., David, D., Bata, J., Prensier, G. & Vivares, C.P. 1994 On small genomes in eukaryotic organisms: molecular karyotypes of two microsporidian species (Protozoa) parasites of vertebrates. Comptes Rendues de l' Academie des Sciences. Paris Life Science 317, 300-404.

    Google Scholar 

  • Champliaud, D., Gobet, P., Naciri, M., Vagner, O., Lopez, J., Buisson, J.C., Varga, I., Harly, G., Mancassola, R. & Bonnin, A. 1998 Failure to di.erentiate Cryptosporidium parvum from C. meleagridis based on PCR amplification of eight DNA sequences. Applied and Environmental Microbiology 64, 1454-1458.

    CAS  Google Scholar 

  • De Jonge, P., DeJpngh, F.C.M., Meijers, R., Steensma, H.Y. & Scheffers, W.A. 1986 Orthogonal-field-alternation gel electrophoresis banding patterns of DNA from yeasts. Yeast 2, 193-204.

    Article  CAS  Google Scholar 

  • Ehrlich, G.D. & Greenberg, S.J. 1994 Detection of hepatitis C virus RNA in liver and plasma by Reverse transriptase PCR using Liquid hybridisation. In: PCR-based Diagnostics in Infectious Diseases, pp. 616. Blackwell Scientific Publications. ISBN 0-86542252-4.

  • Fonstein, M. & Haselkorn, R. 1995 Physical mapping of bacterial genomes. Journal of Bacteriology 177, 3361-3369.

    CAS  Google Scholar 

  • Franzen, C., Kiippers, R., Miiller, A., Salzberger, B., Fatkenheuer, G., Vetten, B., Diehl, V. & Schrappe, M. 1996 Genetic evidence for latent Septata intestinalis infection in Human Immunodeficiency Virus-infected patients with intestinal Microsporidiosis. The Journal of Infectious Diseases 173, 1038-1040.

    CAS  Google Scholar 

  • Giannini, S.H., Schittini, M., Keithly, J.S., Warhurton, P.W., Cantor, C.R. & Van Der Ploeg, L.H.T. 1986 Karyotype analysis of Leishmania species and its use in classification and clinical diagnosis. Science 232, 762-765.

    CAS  Google Scholar 

  • Hartskeerl, R.A., Schuitema, A.R.J. & De Wachter, R. 1993 Secondary structure of the small sub unit ribosomal RNA sequence of the microsporidium Encephalitozoon cuniculi. Nucleic Acids Research 21, 1489.

    CAS  Google Scholar 

  • Hazard, E.I., Andreadis, T.G., Joslyn, D.J. & Ellis, E.A. 1979 Meiosis and its implications in the life cycles of Amblyospora and Parathelohania (Microspora). Journal of Parasitology 65, 117-122.

    Article  Google Scholar 

  • Hirai, H. & Loverde, P.T. 1995 FISH Techniques for construction of Physical Maps on Schistosome Chromosomes. Parasitology Today 11, 310-314.

    Article  CAS  Google Scholar 

  • Irby, W.S., Huang, Y.S., Kawanishi, C.Y. & Brooks, W.M. 1986 Immunoblot analysis of exospore polypeptides from some entomophilic microsporidia. Journal of Protozoology 33, 14-20.

    CAS  Google Scholar 

  • Johnson, D.W., Pieniazek, N.J. & Rose, J.B. 1993 DNA probe hybridization and PCR detection of Cryptosporidium compared to immuno-fluorescence assay. Water Science and Technology 27, 77-84.

    Google Scholar 

  • Johnson, D.W., Pieniazek, N.J., Griffin, D.W., Misener, I. & Rose, J.B. 1995 Development of a PCR protocol for sensitive detection of Cryptosporidium oocysts in water samples. Applied and Environmental Microbiology 61, 3849-3855.

    CAS  Google Scholar 

  • Kariem, A.E.F.M., Washurst, D.C. & McDonald, V. 1994 Detection and species identification of Cryptosporidium oocysts using a system based on PCR and endonuclease restriction. Parasitology 109, 19-22.

    Article  Google Scholar 

  • Kawarabata, T. & Hayasaka, S. 1987 An enzyme-linked immunosorbent assay to detect alkali-soluble spore surface antigens of strains of Nosema bombycis (Microspora: Nosematidae). Journal of Invertebrate Pathology 50, 118-123.

    Article  Google Scholar 

  • Kawakami, Y., Inoue, T., Ito, K., Hanawa, C., Ando, T., Iwano, H. & Ishibara, R. 1994 Identification of a chromosome harboring the small subunit ribosomal RNA gene of Nosema bombycis. Journal of Invertebrate Pathology 64, 147-148.

    Article  CAS  Google Scholar 

  • Malone, L.A. & McIvor, C.A. 1993 PFGE of DNA from 4 microsporidia isolates. Journal of Invertebrate Pathology 61, 243-248.

    Article  Google Scholar 

  • Malone, L.A. & McIvor, C.A. 1995 DNA probes for 2 microsporidia, N. bombycis and N. costelytrae. Journal of Invertebrate Pathology 65, 269-273.

    Article  CAS  Google Scholar 

  • Malone, L.A. & McIvor, C.A. 1996 Use of nucleotide sequence Data to identify a microsporidian pathogen of Pieris rapae (Lepidoptera, Pieridae). Journal of Invertebrate Pathology 68, 231-238.

    Article  CAS  Google Scholar 

  • Malone, L.A., Broadwell, A.H., Lindridge, E.T., McIvor, C.A. & Ninham, J.A. 1994 Ribosomal RNA genes of two microsporidia, Nosemaapis and Vavraia oncoperae, are very variable. Journal of Invertebrate Pathology 64, 151-152.

    Article  CAS  Google Scholar 

  • Munderloh, U.G., Kurtti, T.J. & Ross, S.E. 1990 Electrophoretic characterization of chromosomal DNA from two microsporidia. Journal of Invertebrate Pathology 56, 243-248.

    Article  CAS  Google Scholar 

  • Neefs, J.M., Van de Peer, Y., Goris, A. & De Wachter, R. 1991 Compilation of small ribosomal subunit RNA sequences. Nucleic Acids Research 19, 1987-2015.

    CAS  Google Scholar 

  • Pereira, M.D.G.C., Atwill, E.R., Crawford, M.R. & Lefebvre, R.B. 1998 DNA Sequence Similarity between California Isolates of Cryptosporidium parvum. Applied and Environmental Microbiology 64, 1584-1586.

    CAS  Google Scholar 

  • Pinevich, A., Gromov, B., Mamkaeva, K. & Nasonova, E. 1997 Study of molecular karyotypes in Amoeboaphelidium protococcarum, the endotrophic parasite of chlorophycean alga Scenedesmus. Current Microbiology 34, 122-126.

    Article  CAS  Google Scholar 

  • Rademaker, J.L.W. & Janse, J.D. 1994 Detection and identification of Clavibacter michiganesis subsp. sepedonicus and Clavibacter michiganensis subsp. michiganensis by nonradioactive hybridization, polymerase chain reaction and restriction enzyme analysis. Canadian Journal of Microbiology 40, 1007-1018.

    CAS  Google Scholar 

  • Ralph, D., McClelland, M., Welsh, J., Baranton, G. & Perolat, P. 1993 Leptospira species categorized by arbitrarily primed polymerase chain reaction (PCR) and by mapped restriction polymorphisms in PCR amplified rRNA genes. Journal of Bacteriology 175, 973-981.

    CAS  Google Scholar 

  • Rochelle, P.A., De Leon, R., Stewart, M.H. & Wolfe, R.L. 1997 Comparison of primers and optimization of PCR conditions for detection of Cryptosporidium parvum and Giardia lamblia in water. Applied and Environmental Microbiology 63, 106-114.

    CAS  Google Scholar 

  • Sato, R. & Watanabe, H. 1980 Purification of mature microsporidian spores by isodensity equilibrium centrifugate. Journal of Sericulture Science Japan 49, 512-516.

    Google Scholar 

  • Schwartz, D.C. & Cantor, C.R. 1984 Separation of yeast chromosome sized DNAs by pulsed field gradient gel electrophoresis. Cell 37, 67-75.

    Article  CAS  Google Scholar 

  • Shimizu, S., Nishida, Y., Yoshioka, Y. & Matsumoto, T. 1991 Separation of chromosomal DNA molecules from Paecilomyces fumosoroseus by pulsed field electrophoresis. Journal of Invertebrate Pathology 58, 461-463.

    Article  CAS  Google Scholar 

  • Sironmani, T.A. 1997 Detection of Nosema bombycis infection in the silkworm Bombyx mori by westernblot analysis. Sericologia 37, 209-216.

    Google Scholar 

  • Sironmani, T.A. 1999a Biochemical characterization of the microsporidian Nosema bombycis spore proteins. World Journal of Microbiology & Biotechnology 15, 213-220.

    CAS  Google Scholar 

  • Sironmani, T.A. 1999b Immunological characterization of spore proteins of the microsporidian Nosema bombycis. World Journal of Microbiology & Biotechnology 15, 607-613.

    Article  CAS  Google Scholar 

  • Steffan, R.J. & Atlas, R.M. 1991 Polymerase Chain reaction: applications in environmental microbiology. Annual Review of Microbiology 45, 137-161.

    Article  CAS  Google Scholar 

  • Streett, D.A. & Briggs, J.D. 1982a An evaluation of SDS-PAGE for the identification of microsporidia. Journal of Invertebrate Pathology 40, 159-165.

    Article  CAS  Google Scholar 

  • Streett, D.A. & Briggs, J.D. 1982b Variation in spore polypeptides from 4 sp. of Vairimorpha. Biological Systematic Ecology 10, 161-165.

    Article  CAS  Google Scholar 

  • Tsai, Y.L., Palmer, C.J. & Sangermano, L.R. 1993 Detection of Escherichia coli in sewage and sludge by polymerase chain reaction. Applied and Environmental Microbiology 59, 353-357.

    CAS  Google Scholar 

  • Undeen, A.H. & Cookborn, A.F. 1989 The extraction of DNA from microsporidia spores. Journal of Invertebrate Pathology 54, 132-133.

    Article  Google Scholar 

  • Vavra, J. 1965 Etude au microscope electronique de la morphologie et du development de quelques microsporidies. Comptes Rendues de l' Academie des Sciences Paris 261, 3467-3470.

    CAS  Google Scholar 

  • Van Keulen, H., Campbell, S.R. & Jarroll, E.L. 1989 Differentation between Giardia lamblia and Giardia muris by southern blot analysis and orthogonal-field-alternation gel electrophoresis karyotype analysis. In: Biochemistry and Molecular Biology of Anaerobic Protozoa, eds. Lloyd, D., Coombs, G.H. & Paget, T.A. pp. 217-231. Chur: Harwood Academic. ISBN 3-71864943-8.

    Google Scholar 

  • Vossbrinck, C.R., Maddox, J.V., Friedman, B.A., Debrunner Vossbrinck, B.A. & Woese, C.R. 1987 Ribosomal RNA sequences suggests microsporidia are extremely ancient eukaryotes. Nature (London) 326, 411-414.

    Article  CAS  Google Scholar 

  • Vossbrinck, C.R., Baker, M.D., Didier, E.S., Debrunner-Vossbrinck, B.A. & Shadduck, J.A. 1993 Ribosomal DNA sequences of Encephalitozoon hellem and Encephalitozoon cuniculi: Species identification and phylogenetic construction. Journal of Eukaryotic Microbiology 40, 354-362.

    CAS  Google Scholar 

  • Weiss, J.B., Van Keulen, H. & Nash, T.E. 1992 Classification of subgroups of Giardia lamblia based upon ribosomal RNA gene sequence using the polymerase chain reaction. Molecular Biochemistry and Parasitology 54, 73.

    Article  CAS  Google Scholar 

  • Welsh, J., Petersen, C. & Mc Clelland, M. 1991 Polymorphisms generated by arbitrarily primed PCR in the mouse; Application to stain identification and genetic mapping. Nucleic Acids Research 19, 303-306.

    CAS  Google Scholar 

  • Welsh, J., Pretzman, C., Postic, D., Saintgirons, I., Baranton, G. & Mc Clelland, M. 1992 Genomic fingerprinting by arbitrarily primed polymerase chain reaction resolves Borrelia burgdorferi in to three distinct phyletic groups. International Journal of Systematic Bacteriology 42, 370-377.

    Article  CAS  Google Scholar 

  • Yamaichi, Y., Lida, T., Sam park, K. & Honda, T. 1999 Physical and genetic map of the genome of Vibrio parahaemolyticus: presence of two chromosomes in Vibrio species. Molecular Microbiology 31, 1513-1521.

    Article  CAS  Google Scholar 

  • Zhu, X., Wittner, M., Tanowitz, H.B., Cali, A. & Weiss, L.M. 1993a Nucleotide sequence of the small ribosomal RNA of Encephalitozoon cuniculi. Nucleic Acids Research 21, 1315.

    CAS  Google Scholar 

  • Zhu, X., Wittner, M., Tanowitz, H.B., Cali, A. & Weiss, L.M. 1993b Small subunit rRNA Sequence of Enterocytozoon bieneusi and its potential diagnostic role with use of the polymerase chain reaction. Journal of infectious Diseases 168, 1570-1575.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sironmani, T. Preliminary genomic characterization of microsporidian Nosema bombycis. World Journal of Microbiology and Biotechnology 16, 533–540 (2000). https://doi.org/10.1023/A:1008986614938

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1008986614938

Navigation