Hostname: page-component-7c8c6479df-94d59 Total loading time: 0 Render date: 2024-03-27T16:40:50.528Z Has data issue: false hasContentIssue false

Identification and characterization of three Encephalitozoon cuniculi strains

Published online by Cambridge University Press:  06 April 2009

E. S. Didier
Affiliation:
Department of Microbiology, Tulane Regional Primate Research Center, 18703 Three Rivers Road, Covington, LA 70433, USA
C. R. Vossbrinck
Affiliation:
Office of Agricultural Entomology, University of Illinois, Urbana, IL 61801, USA
M. D. Baker
Affiliation:
Office of Agricultural Entomology, University of Illinois, Urbana, IL 61801, USA
L. B. Rogers
Affiliation:
Department of Microbiology, Tulane Regional Primate Research Center, 18703 Three Rivers Road, Covington, LA 70433, USA
D. C. Bertucci
Affiliation:
Department of Microbiology, Tulane Regional Primate Research Center, 18703 Three Rivers Road, Covington, LA 70433, USA
J. A. Shadduck
Affiliation:
Department of Veterinary Pathobiology, Texas A & M University, College Station, TX 77843, USA

Summary

Microsporidia are increasingly recognized as causing opportunistic infections in immunocompromised individuals. Encephalitozoon cuniculi is probably the most studied mammalian microsporidian that infects insects and mammals, including man. In this study, 8 E. cuniculi isolates were compared and were found to fall into 3 strains. Strain type I includes the rabbit type isolate, as well as isolates from an additional rabbit, a dwarf rabbit, and a mouse. Strain type II includes 2 murine isolates and strain type III includes 2 isolates obtained from domestic dogs. By SDS-PAGE, the 3 strains differ primarily in the molecular weight range of 54–59 kDa where strain type I displays an apparent broad singlet at 57 kDa, strain type II displays an apparent doublet at 54 and 58 kDa, and strain type III displays an apparent broad band at 59 kDa. Antigenic differences were detected in the molecular weight regions of 54–58 kDa as well as 28–40 kDa by Western blot immunodetection using murine antisera raised against E. cuniculi, Encephalitozoon hellem, and the Encephalitozoon-like Septata intestinalis. Polymerase chain reaction (PCR) products containing only small subunit rDNA sequences from the different E. cuniculi isolates formed homoduplexes whereas PCR products containing intergenic rRNA gene sequences formed heteroduplexes in mobility shift analyses. Fok I digestion of the PCR products containing the intergenic rRNA gene region resulted in unique restriction fragment length polymorphism patterns, and DNA sequencing demonstrated that in the intergenic spacer region, the sequence 5'-GTTT-3' was repeated 3 times in strain type I, twice in strain type II, and 4 times in strain type III. This study indicates that there exist at least 3 E. cuniculi strains which may become important in the epidemiology of human E. cuniculi infections. Furthermore, as additional E. cuniculi isolates are characterized, these strains will be named or reclassified once the criteria for taxonomy and phylogenetic tree construction for microsporidia become better defined.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1995

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Aldras, A. M., Orenstein, J. M., Kotler, D. P., Shadduck, J. A. & Didier, E. S. (1993). Detection of microsporidia by indirect immunofluorescence antibody test using polyclonal and monoclonal antibodies. Journal of Clinical Microbiology 32, 608–12.Google Scholar
Bergquist, N. R., Morfeldt-Mansson, L., Pehrson, P. O., Petrini, B. & Wasserman, J. (1984 a). Antibody against Encephalitozoon cuniculi in Swedish homosexual men. Scandinavian Journal of Infectious Diseases 16, 389–91.Google Scholar
Bergquist, N. R., Stintzing, G., Smedman, L., Waller, T. & Anderson, T. (1984 b). Diagnosis of encephalitozoonosis in man by serological tests. British Medical Journal 288, 902.Google Scholar
Blake, M. S., Johnston, K. H., Russell-Jones, G. J. & Gottschlich, E. C. (1984). A rapid, sensitive method for detection of alkaline-phosphatase-conjugated antibody on Western blots. Analytical Biochemistry 136, 175–9.Google Scholar
Botha, W. S., Van Dellen, A. F. & Stewart, C. G. (1979). Canine encephalitozoonosis in South Africa. Journal of the South African Veterinary Medical Association 5, 135–44.Google Scholar
Bryan, R. T., Cali, A., Owen, R. L. & Spencer, H. C. (1990). Microsporidia. Opportunistic pathogens in patients with AIDS. Progress in Clinical Parasitology 2, 126.Google Scholar
Cali, A., Kotler, D. P. & Orenstein, J. M. (1993). Septata intestinalis n.g., n.sp., an intestinalis microsporidian associated with chronic diarrhea and dissemination in AIDS patients. Journal of Protozoology 40, 101–12.Google Scholar
Cali, A., Weiss, L. M., Takvorian, P., Tanowitz, P. & Wittner, M. (1994). Ultrastructural identification of AIDS associated microsporidiosis. Journal of Eukaryotic Microbiology 41, 24S.Google Scholar
Canning, E. U. & Hollister, W. S. (1992). Human infections with microsporidia. Reviews in Medical Microbiology 3, 3542.Google Scholar
Canning, E. U. & Lom, J. (1986). The Microsporidia of Vertebrates. New York: Academic Press.Google Scholar
Chupp, G. L., Alroy, J., Adelman, L. S., Breen, J. C. & Skolnick, P. R. (1993). Myositis due to Pleistophora (Microsporidia) in a patient with AIDS. Clinical Infectious Diseases 16, 1521.Google Scholar
Cox, J. C. & Pye, D. (1975). Serodiagnosis of nosematosis by immunofluorescence using cell culture grown organisms. Laboratory Animals 9, 297304.Google Scholar
Davis, R. M., Font, R. L., Keisler, M. S. & Shadduck, J. A. (1990). Corneal microsporidiosis. A case report including ultrastructural observations. Ophthalmology 97, 953–7.Google Scholar
De Groote, M. A., Visvesvara, G. S., Wilson, M. L., Pieniazek, N. J., Slemenda, S. B., Dasilva, A. J., Leitch, G. J., Bryan, R. T. & Reves, R. (1995). Polymerase chain reaction and culture confirmation of disseminated Encephalitozoon cuniculi in a patient with AIDS: Successful therapy with albendazole. Journal of Infectious Disease 171, 1375–8.Google Scholar
Desportes, I., Charpentier, Y. Le, Galian, A., Bernard, F., Cochand-Priollet, B., Lavergne, A., Ravisse, P. & Modigliani, R. (1985). Occurrence of a new microsporidian, Enterocytozoon bieneusi n.g., n.sp., in the enterocytes of a human patient with AIDS. Journal of Protozoology 32, 250–4.Google Scholar
Didier, E. S., Didier, P. J., Friedberg, D. N., Stenson, S. M., Orenstein, J. M., Yee, R. W., Tio, F. O., Davis, R. M., Vossbrinck, C. R., Millichamp, N. & Shadduck, J. A. (1991 a). Isolation and characterization of a new human microsporidian, Encephalitozoon hellem (n.sp.), from three AIDS patients with keratocon-junctivitis. Journal of Infectious Diseases 163, 617–21.Google Scholar
Didier, P. J., Orenstein, J. M., Didier, E. S. & Shadduck, J. A. (1991 b). Fine structure of a new human microsporidian, Encephalitozoon hellem, in culture. Journal of Protozoology 38, 502–7.Google Scholar
Friedberg, D. N., Stenson, S. M., Orenstein, J. M., Tierno, P. & Chales, N. (1990). Microsporidial keratoconjunctivitis in the acquired immunodeficiency syndrome. Archives in Ophthalmology 108, 504–8.Google Scholar
Hollister, W. S., Canning, E. U. & Colbourn, N. I. (1994). A species of Encephalitozoon isolated from an AIDS patient: criteria for species differentiation. Folia Parasitologica 40, 293–5.Google Scholar
Hollister, W. S., Canning, E. U., Colbourn, N. I. & Aarons, E. J. (1995). Encephalitozoon cuniculi isolated from the urine of an AIDS patient, which differs from a canine isolate. Journal of Eukaryotic Microbiology (In the press).Google Scholar
Hollister, W. S., Canning, E. U., Colbourn, N. I., Curry, A. & Lacey, C. J. N. (1993). Characterization of Encephalitozoon hellem (Microspora) isolated from the nasal mucosa of a patient with AIDS. Parasitology 107, 351–8.Google Scholar
Kasper, L. H. & Boothroyd, J. C. (1993). Toxoplasma gondii and toxoplasmosis. In Immunology and Molecular Biology of Parasitic Infections, 3rd Edn (ed. Warren, K. S.), Oxford: Blackwell Scientific Publications.Google Scholar
Ledford, D. K., Overman, M. D., Gonzalo, A., Cali, A., Mester, W. & Lockey, R. F. (1985). Microsporidiosis myositis in a patient with acquired immunodeficiency syndrome. Annals in Internal Medicine 102, 628–30.Google Scholar
Levaditi, C., Nicolau, S. & Schoen, R. (1924). L'étiogie de l'encéphalite épizootique du lapin, dans ses rapports avec l'étude expérimentale de l'encéphalite léthargique Encephalitozoon cuniculi (nov. spec.). Annales de I' Institute Pasteur, (Paris) 38, 651711.Google Scholar
Margileth, A. M., Strand, A. J., Chandra, R., Neafie, R., Blum, M. & McCully, R. M. (1973). Disseminated nosematosis in an immunologically compromised infant. Archives in Pathology 95, 341–3.Google Scholar
Montrey, R. D., Shadduck, J. A. & Pakes, S. P. (1973). In vitro host range of three isolates of Encephalitozoon (Nosema). Journal of Infectious Diseases 98, 306–11.Google Scholar
Nordstoga, K. & Westbye, K. (1976). Polyarteritis nodosa associated with nosematosis in blue foxes. Acta Pathologica et Microbiologica Scandinavica, Section A 84, 291–6.Google Scholar
Orenstein, J. M. (1991). Microsporidiosis in the acquired immunodeficiency syndrome. Journal of Parasitology 77, 843–64.Google Scholar
Orenstein, J. M., Tenner, M., Cali, A. & Kotler, D. P. (1992). A microsporidian previously undescribed in humans, infecting enterocytes and macrophages, and associated with diarrhea in an acquired immunodeficiency syndrome patient. Human Pathology 23, 722–8.Google Scholar
Pakes, S. P., Shadduck, J. A. & Cali, A. (1975). Fine structure of Encephalitozoon cuniculi from rabbits, mice, and hamsters. Journal of Protozoology 22, 481–8.Google Scholar
Pang, V. F. & Shadduck, J. A. (1985). Susceptibility of cats, sheep, and swine to a rabbit isolate of Encephalitozoon cuniculi. American Journal of Veterinary Research 46, 1071–7.Google Scholar
Pattison, M., Clegg, F. G. & Duncan, A. L. (1971). An outbreak of encephalitomyelitis in broiler rabbits caused by Nosema cuniculi. Veterinary Record 88, 404–5.Google Scholar
Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: A Laboratory manual, 2nd EdnCold Spring Harbor, New York: Cold Spring Harbor Laboratory Press.Google Scholar
Schmidt, E. C. & Shadduck, J. A. (1983). Murine encephalitozoonosis model for studying the host-parasite relationship of a chronic infection. Infection and Immunity 40, 936–42.Google Scholar
Schmidt, E. C. & Shadduck, J. A. (1984). Mechanisms of resistance to the intracellular protozoan Encephalitozoon cuniculi in mice. Journal of Immunology 133, 2712–19.Google Scholar
Schuitema, A. R. J., Hartskeerl, R. A., Gool, T. Van, Laxminarayan, R. & Terpstra, W. J. (1993). Application of the polymerase chain reaction for the diagnosis of microsporidiosis. AIDS 7, S57–S61.Google Scholar
Schwartzman, J. D., Boothroyd, J. C. & Kasper, L. H. (1994). Toxoplasma workshop overview. Journal of Eukaryotic Microbiology 41, 19S21S.Google Scholar
Shadduck, J. A. (1969). Nosema cuniculi: in vitro isolation. Science 166, 516–17.Google Scholar
Shadduck, J. A. (1989). Human microsporidiosis in AIDS. Reviews in Infectious Diseases 11, 203–7.Google Scholar
Shadduck, J. A. & Baskin, G. (1989). Serologic evidence of Encephalitozoon cuniculi infection in a colony of squirrel monkeys (Saimiri sciureus). Laboratory Animal Science 39, 328–30.Google Scholar
Shadduck, J. A. & Greeley, E. (1989). Microsporidia and human infections. Clinical Microbiology Reviews 2, 158–65.CrossRefGoogle ScholarPubMed
Shadduck, J. A. & Pakes, S. P. (1971). Spontaneous diseases of laboratory animals which interfere with biomedical research. Encephalitozoonosis and toxoplasmosis. American Journal of Pathology 64, 657–74.Google Scholar
Shadduck, J. A., Bendele, R. & Robinson, G. T. (1978). Isolation of the causative organism of canine encephalitozoonosis. Veterinary Pathology 15, 449–60.Google Scholar
Shadduck, J. A., Meccoli, R. A., Davis, R. & Font, R. L. (1990). First isolation of a microsporidian from a human patient. Journal of Infectious Diseases 162, 773–6.CrossRefGoogle Scholar
Shadduck, J. A., Watson, W. T., Pakes, S. P. & Cali, A. (1979). Animal infectivity of Encephalitozoon cuniculi. Journal of Parasitology 65, 123–9.Google Scholar
Soto, D. & Sukumar, S. (1992). Improved detection of mutations in the p53 gene in human tumors as single-stranded conformation polymorphs and double-stranded heteroduplex DNA. PCR Methods and Applications 2, 96–8.Google Scholar
Sprague, V. (1974). Nosema connori n.sp., microsporidian parasite of man. Transactions of the American Microscopy Society 93, 400–3.Google Scholar
Sprague, V., Becnel, J. J. & Hazard, E. I. (1992). Taxonomy of phylum Microspora. Critical Reviews in Microbiology 18, 285395.Google Scholar
Stewart, C. G., Dellen, A. F. Van & Botha, W. S. (1979). Canine encephalitozoonosis in kennels and the isolation of Encephalitozoon cuniculi in cell culture. Journal of South African Veterinary Association 50, 165–8.Google Scholar
Terada, S., Reddy, R. K., Jeffers, L. J. & Cali, A. (1987). Microsporidian hepatitis in the acquired immunodeficiency syndrome. Annals in Internal Medicine 107, 61–2.CrossRefGoogle ScholarPubMed
Towbin, H., Staehelin, T. & Gordon, J. (1984). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. Procedures and some applications. Proceedings of the National Academy of Sciences, USA 76, 4350–4.Google Scholar
Dellen, A. F. Van, Stewart, C. G. & Botha, W. S. (1989). Studies of encephalitozoonosis in vervet monkeys (Cercopithecus pygerythrus) orally inoculated with spores of Encephalitozoon cuniculi isolated from dogs (Canis familiaris). Onderstepoort Journal of Veterinary Research 56, 122.Google Scholar
Gool, T. Van, Canning, E. U., Gilis, H., Van Den Bergh Weerman, M. A., Eeftinck, Schattenkerk J. K. M. & Dankert, J. (1994). Septata intestinalis frequently isolated from stool of AIDS patients with a new cultivation method. Parasitology 109, 281–9.Google Scholar
Vávra, J., Bedrník, P. & Činatl, J. (1972). Isolation and in vitro cultivation of the mammalian microsporidia Encephalitozoon cuniculi. Folia Parasitologica 19, 349–54.Google Scholar
Visvesvara, G. S., Dasilva, A. J., Croppo, G. P., Pieniazek, N. J., Slemenda, S., Leitch, G. J., Ferguson, D., Wallace, S., Tyrrel, I. & Medor, J. (1994). Continuous cultivation, serological and molecular characterization of Septata intestinalis from an AIDS patient with disseminated microsporidosis. Society of Protozoologists 47th Annual Meeting C4, (Abstract).Google Scholar
Vossbrinck, C. R. & Woese, C. R. (1986). Eukaryotic ribosomes that lack a 5.8S RNA. Nature, London 320, 287–8.Google Scholar
Vossbrinck, C. R., Baker, M. D., Didier, E. S., Debrunner, Vossbrinck B. A. & Shadduck, J. A. (1993). rDNA sequences of Encephalitozoon hellem and Encephalitozoon cuniculi: species identification and phylogenetic construction. Journal of Eukaryotic Microbiology 40, 354–62.Google Scholar
Vossbrinck, C. R., Maddox, J. V., Frieman, S., Debrunner-Vossbrinck, B. A. & Woese, C. R. (1987). Ribosomal RNA sequences suggest microsporidia are extremely ancient eukaryotes. Nature, London 326, 411–14.Google Scholar
Weber, R., Bryan, R. T., Schwartz, D. A. & Owen, R. L. (1994). Human microsporidial infections. Clinical Microbiology Reviews 7, 426–61.Google Scholar
Weiser, J. (1964). On the taxonomic position of the genus Encephalitozoon Levaditi, Nicolau & Schoen, 1923 (Protozoa: Microsporidia). Parasitology 54, 749–51.Google Scholar
Weiser, J. (1965). Microsporidian infections of mammals and the genus Encephalitozoon. International Congress in Parasitology 1, 445–6.Google Scholar
Wright, J. H. & Craighead, E. M. (1922). Infectious motor paralysis in young rabbits. Journal of Experimental Medicine 36, 135–40.Google Scholar
Yee, R. W., Tio, F. O., Martinez, J. A., Held, K. S., Shadduck, J. A. & Didier, E. S. (1991). Resolution of microsporidial epithelial keratopathy in a patient with AIDS. Ophthalmology 98, 196201.Google Scholar
Zeman, D. H. & Baskin, G. B. (1985). Encephalitozoonosis in squirrel monkeys (Saimiri sciureiis). Veterinary Pathology 22, 2431.Google Scholar
Zender, H. O., Arrigoni, E., Eckert, J. & Kapanci, Y. (1989). A case of Encephalitozoon cunicitli peritonitis in a patient with AIDS. American Journal of Clinical Pathology 92, 352–6.Google Scholar