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Volume 150, Issue 5

Spore morphotypes of (microsporidia) described microscopically and confirmed by PCR of individual spores microdissected from smears by position ablative laser microbeam microscopy Free

Abstract

Development of , a parasite of the red imported fire ant (), until recently was thought to include formation of two types of spores: unicellular meiospores, maturing inside sporophorous vesicles in sets of eight (octospores); and -like binuclear free spores. Megaspores, discovered in 2001, develop primarily in alates and are morphologically distinct from the two previously known types of spores. The role of megaspores in the life cycle, as well as their existence, has been questioned. The current research includes light and electron microscopic descriptions of the three major spore morphotypes characteristic of development. In addition, individual octospores and megaspores were isolated into groups of 8–20 from methanol-fixed and Calcofluor-stained smears of the infected ants for subsequent PCR analysis by the laser pressure catapulting function of a position ablative laser microbeam microscope, a technique applied for the first time to research of microsporidia. The PCR-amplified SSU rDNA nucleotide sequences from octospores and megaspores were identical. This, along with the consistency with which megaspores are detected in infected ants, demonstrates that megaspores are integral to the life cycle of .

  • Received:
  • Accepted:
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  • Published Online:
Keyword(s): LPC, laser pressure catapulting and PALM, position ablative laser microbeam
SGM
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2004-05-01
2024-05-10
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References

  1. Banks W. A., Lofgren C. S., Jouvenaz D. P., Stringer C. E., Bishop P. M., Williams D. F., Wojcik D. P., Glancey B. M. 1981 Techniques for Collecting, Rearing, and Handling Imported Fire Ants US Department of Agriculture Science and Education Administration Publications, AAT; NS-21
  2. Becnel J. J., Andreadis T. G. 1999; Microsporidia in insects. In The Microsporidia and Microsporidiosis pp. 447–501Edited by Wittner M., Weiss L. M. Washington, DC: American Society for Microbiology;
     [Google Scholar]
  3. Becnel J. J., Sprague V., Fukuda T., Hazard E. I. 1989; Development of Edhazardia aedis (Kudo, 1930) n.g., n.comb. (Microsporidia: Amblyosporidae) in the mosquito Aedes aegypti (L.) (Diptera: Culicidae). J Protozool 36:119–130 [CrossRef]
     [Google Scholar]
  4. Cao Z., Wanagat J., McKiernan S., Aiken J. M. 2001; Mitochondrial DNA deletion mutations are concomitant with ragged red regions of individual, aged muscle fibers: analysis by laser-capture microdissection. Nucleic Acids Res 29:4502–4508 [CrossRef]
     [Google Scholar]
  5. Cook T. E. 2002; Studies of naturally occurring Thelohania solenopsae(Microsporidia: Thelohaniidae) infection in red imported fire ants, Solenopsis invicta (Hymenoptera: Formicidae). Environ Entomol 31:1091–1096 [CrossRef]
     [Google Scholar]
  6. Cor A., Vogt N., Malfoy B. 2002; Microdissection for cancer analysis. Folia Biol 48:3–8
     [Google Scholar]
  7. Didier E. S., Orenstein J. M., Aldras A. A., Bertucci D. L., Rogers B., Janney F. A. 1995; Comparison of three staining methods for detecting microsporidia in fluids. J Clin Microbiol 33:3138–3145
     [Google Scholar]
  8. Eltoum I. A., Siegal G. P., Frost A. R. 2002; Microdissection of histologic sections: past, present, and future. Adv Anat Pathol 9:316–322 [CrossRef]
     [Google Scholar]
  9. Fend F., Kremer M., Quintanilla-Martinez L. 2000; Laser capture microdissection: methodical aspects and applications with emphasis on immuno-laser capture microdissection. Pathobiology 68:209–214 [CrossRef]
     [Google Scholar]
  10. Grant K., Jerome W. G. 2002; Laser capture microdissection as an aid to ultrastructural analysis. Microsc Microanal 8:170–175 [CrossRef]
     [Google Scholar]
  11. Jouvenaz D. P., Ellis E. A. 1986; Vairimorpha invictae n.sp.(Microspora: Microsporidia), a parasite of the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae). J Protozool 33:24–29
     [Google Scholar]
  12. Jouvenaz D. P., Hazard E. I. 1978; New family, genus, and species of microsporidia (Protozoa: Microsporida) from the tropical fire ant, Solenopsis geminata (Fabricius) (Insecta: Formicidae). J Protozool 25:24–29 [CrossRef]
     [Google Scholar]
  13. Knell J. D., Allen G. E., Hazard E. I. 1977; Light and electron microscope study of Thelohania solenopsae n.sp.(Microsporida: Protozoa) in the red imported fire ant, Solenopsia invicta. J Invertebr Pathol 29:192–200 [CrossRef]
     [Google Scholar]
  14. Lehmann U., Glockner S., Kleeberger W., Feist H., von Wasielewski R., Kreipe H. 2000; Detection of gene amplification in archival breast cancer specimens by laser-assisted microdissection and quantitative real-time polymerase chain reaction. Am J Pathol 156:1855–1864 [CrossRef]
     [Google Scholar]
  15. Maddox J. V., Baker M. D., Jeffords M. R., Kuras M., Linde A., Solter L. E., McManus M. L., Vavra J., Vossbrinck C. R. 1999; Nosema portugal, n.sp., isolated from gypsy moths (Lymantria dispar L.) collected in Portugal. J Invertebr Pathol 73:192–200
     [Google Scholar]
  16. MetaView 1998 Meta Imaging Series 4.5 West Chester, PA: Universal Imaging Corporation;
  17. Milks M. L., Sokolova Y. Y., Isakova I. A., Fuxa J. R., Mitchell F., Snowden K. F., Vinson S. B. 2004; Comparative effectiveness of light-microscopic techniques and PCR in detecting Thelohania solenopsae(Microsporidia) infections in red imported fire ants (Solenopsis invicta). J Eukaryot Microbiol 51:187–191 [CrossRef]
     [Google Scholar]
  18. Moser B. A. 1995 Comparative analysis of microsporidia of fire ants Solenopsis richteri and S. invicta PhD dissertation, University of Florida; Gainesville, USA:
  19. Moser B. A., Becnel J. J., Maruniak J., Patterson R. S. 1998; Analysis of the ribosomal DNA sequences of the microsporidia Thelohania and Vairimorpha of fire ants. J Invertebr Pathol 72:154–159 [CrossRef]
     [Google Scholar]
  20. Moser B. A., Becnel J. J., Williams D. F. 2000; Morphological and molecular characterization of the Thelohania solenopsae complex (Microsporidia: Thelohaniidae). J Invertebr Pathol 75:174–177 [CrossRef]
     [Google Scholar]
  21. Obiakor H., Sehgal D., Dasso J. F., Bonner R. F., Malekafzali A., Mage R. G. 2002; A comparison of hydraulic and laser capture microdissection methods for collection of single B cells, PCR, and sequencing of antibody VDJ. Anal Biochem 306:55–62 [CrossRef]
     [Google Scholar]
  22. Oi D. H., Williams D. F. 2002; Impact of Thelohania solenopsae(Microsporidia: Thelohaniidae) on polygyne colonies of red imported fire ants (Hymenoptera: Formicidae). J Econ Entomol 95:558–562 [CrossRef]
     [Google Scholar]
  23. Oi D. H., Becnel J. J., Williams D. F. 2001; Evidence of intracolony transmission of Thelohania solenopsae (Microsporidia: Thelohaniidae) in red imported fire ants (Hymenoptera: Formicidae) and first report on spores from pupae. J Invertebr Pathol 78:128–134 [CrossRef]
     [Google Scholar]
  24. Ryan P., Bennett M. W., Aarons S., Lee G., Collins J. K., O'Sullivan G. C., O'Connel J., Shanahan F. 2002; PCR detection of Mycobacterium paratuberculosis in Crohn's disease granulomas isolated by laser capture microdissection. Gut 55:665–670
     [Google Scholar]
  25. Shapiro A. M., Becnel J. J., Oi D. F., Williams D. F. 2003; Ultrastructural characterization and further transmission studies of Thelohania solenopsae from Solenopsis invicta. J Invertebr Pathol 83:177–180 [CrossRef]
     [Google Scholar]
  26. Sokolova Y. Y., Fuxa J. R. 2001; Development of Thelohania solenopsae in red imported fire ants Solenopsis invicta from polygynous colonies results in formation of three spore types. J Eukaryot MicrobiolSuppl85S
     [Google Scholar]
  27. Stark R., Rubio-Sierra F. J., Thalhammer S., Heckl W. M. 2003; Combined nanomanipulation by atomic force microscopy and UV-laser ablation for chromosomal dissection. Eur Biophys J Biophys Lett 32:33–39
     [Google Scholar]
  28. STATISTICA 1996 STATISTICA for Windows release 5.1 StatSoft Inc;1994–1996 Tulsa, OK, USA:
  29. Undeen A. H. 1997; Microsporidia (Protozoa): a Handbook of Biology and Research Techniques. Southern Cooperative Series Bulletin no. 387.: http://pearl.agcomm.okstate.edu/scsb387/content.htm
     [Google Scholar]
  30. Valles S. M., Oi D. H., Perera O. P., Williams D. F. 2002; Detection of Thelohania solenopsae(Microsporidia: Thelohaniidae) in Solenopsis invicta (Hymenoptera: Formicidae) by multiplex PCR. J Invertebr Pathol 81:196–201 [CrossRef]
     [Google Scholar]
  31. Weber R., Bryan R. T., Owen R. L., Wilcox C. M., Gorelkin L., Visvesvara G. 1992; Improved light-microscopic detection of Microsporida spores in stool and duodenal aspirates. New Engl J Med 4:161–166
     [Google Scholar]
  32. Williams D. F., Knue G. J., Becnel J. J. 1998; Discovery of Thelohania solenopsae from the red imported fire ant,Solenopsis invicta, in the United States. J Invertebr Pathol 71:175–176 [CrossRef]
     [Google Scholar]
  33. Williams D. F., Oi D. H., Knue G. J. 1999; Infection of red imported fire ant (Hymenoptera: Formicidae) colonies with the entomopathogen Thelohania solenopsae (Microsporidia: Thelohaniidae). J Econ Entomol 92:830–836 [CrossRef]
     [Google Scholar]
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Spore morphotypes of Thelohania solenopsae (microsporidia) described microscopically and confirmed by PCR of individual spores microdissected from smears by position ablative laser microbeam microscopy
Microbiology 150, 1261 (2004); https://doi.org/10.1099/mic.0.26838-0
/content/journal/micro/10.1099/mic.0.26838-0
/content/journal/micro/10.1099/mic.0.26838-0
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