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Evidence of infection in tortoises by Chlamydia-like organisms that are genetically distinct from known Chlamydiaceae species


Nasal lavage fluid was collected from 155 tortoises, mostly Testudo spp., that were kept as companion animals and suffered from nasal discharge. Examination for chlamydial DNA by PCR assays targeting the ompA, ompB, and groESL genes, as well as the 16S rRNA signature region and the 16S-23S intergenic spacer, respectively, revealed 16 (10.3%) positive animals. Sequence analysis of PCR products indicated high homology to the family Chlamydiaceae. Phylogenetic trees constructed from partial sequences of the ompA and 16S rRNA genes showed that the present samples clustered outside the nine species of Chlamydia and Chlamydophila. Sequences of the nearest relative, Chlamydophila pecorum, were still clearly distinct from those of the positive tortoise samples. This suggests that the tortoises had been infected by Chlamydia-like agents, the taxonomic identity and pathogenic importance of which has yet to be established.

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  1. Bodetti, T.J., Jacobson, E., Wan, C., Hafner, L., Pospischil, A., Rose, K. and Timms, P., 2002. Molecular evidence to support the expansion of the host range of Chlamydophila pneumoniae to include reptiles as well as humans, horses, koalas and amphibians. Systematic and Applied Microbiology, 25, 146–152

  2. Brown, D.R., Crenshaw, B.C., McLaughlin, G.S., Schumacher, I.M., McKenna, C.E., Klein, P.A., Jacobson, E.R. and Brown, M.B., 1995. Taxonomic analysis of the tortoise mycoplasmas Mycoplasma agassizii and Mycoplasma testudinis by 16S rRNA gene sequence comparison. International Journal of Systematic Bacteriology, 45, 348–350

  3. Brown, M.B., Brown, D.R., Klein, P.A., McLaughlin, P.S., Schumacher, I.M., Jacobson, E.R., Adams, H.P. and Tully, J.G., 2001. Mycoplasma agassizii sp. nov. isolated from the upper respiratory tract of the desert tortoise (Gopherus agassizii) and the gopher tortoise (Gopherus polyphemus). International Journal of Systematic and Evolutionary Microbiology, 51, 413–418

  4. Everett, K.D.E., Bush, R.M. and Andersen, A.A., 1999. Emended description of the order Chlamydiales, proposal of Parachlamydiaceae fam. nov. and Simkaniaceae fam. nov., each containing one monotypic genus, revised taxonomy of the family Chlamydiaceae, including a new genus and five new species, and standards for the identification of organisms. International Journal of Systematic Bacteriology, 49, 415–440

  5. Everett, K.D., 2000. Chlamydia and Chlamydiales: more than meets the eye. Veterinary Microbiology, 75, 109–126

  6. Felsenstein, J., 1993. PHYLIP: Phylogeny Inference Package (Version 3.6). Department of Genetics, University of Washington, Seattle, WA, USA;

  7. Glassick, T., Giffard, P. and Timms, P., 1996. Outer membrane protein 2 gene sequences indicate that Chlamydia pecorum and Chlamydia pneumoniae cause infections in koalas. Systematic and Applied Microbiology, 19, 457–464

  8. Heldstab, A., Berger, R. and Bestetti, G., 1989. Demonstration of virus, chlamydiae and cryptosporidiae in snake gastroenteritis. Herpetopathologia, 1, 101–107

  9. Homer, B.L., Jacobson, E.R., Schumacher, J. and Scherba, G., 1994. Chlamydiosis in mariculture-reared green sea turtles (Chelonia mydas). Veterinary Pathology, 31, 1–7

  10. Hotzel, H., Grossmann, E., Mutschmann, F. and Sachse, K., 2001. Genetic characterization of a Chlamydophila pneumoniae isolate from an African frog and comparison to currently accepted biovars. Systematic and Applied Microbiology, 24, 63–66

  11. Huchzermeyer, F.W., Gerdes, G.H., Foggin, C.M., Huchzermeyer, K.D.A. and Limper, L.C., 1994. Hepatitis in farmed hatchling nile crocodiles (Crocodylus niloticus) due to chlamydial infection. Journal of the South African Veterinary Association, 65, 20–22

  12. Jacobson, E.R., Gaskin, J.M. and Mansell, J., 1989. Chlamydial infection in puff adders (Bitis arietans). Journal of Zoo and Wildlife Medicine, 20, 364–369

  13. Jacobson, E.R. and Telford, S.R., 1990. Chlamydial and poxvirus infections of circulating monocytes of a flap-necked chameleon (Chamaeleo dilepis). Journal of Wildlife Diseases, 26, 572–577

  14. Jacobson, E.R., Heard, D.J., Goebel, T., Bodetti, T. and Timms, P., 2001. Chronic tracheitis, pneumonia and thromboembolic disease in captive Burmese pythons (Python molurus bivittatus), Proceedings of the Association of Reptilian and Amphibian Veterinarians, Orlando, Florida, 119–120

  15. Jacobson, E.R., Origgi, F., Heard, D. and Detrisac, C., 2002. Immunohistochemical staining of chlamydial antigen in emerald tree boas (Corallus caninus). Journal of Veterinary Diagnostic Investigation, 14, 487–494

  16. Jacobson, E.R., Heard, D. and Andersen A., 2004. Identification of Chlamydophila pneumoniae in an emerald tree boa, Corallus caninus. Journal of Veterinary Diagnostic Investigation, 16, 153–154

  17. Kaltenböck, B., Schmeer, N. and Schneider, R., 1997. Evidence for numerous omp1 alleles of porcine Chlamydia trachomatis and novel chlamydial species obtained by PCR. Journal of Clinical Microbiology, 35, 1835–1841

  18. McLaughlin, G.S., 1997. Upper respiratory tract disease in gopher tortoises Gopherus polyphemus: pathology, immune responses, transmission, and implications for conservation and management, (PhD dissertation, University of Florida), 22–24

  19. Sachse, K. and Hotzel, H., 2002. Detection and differentiation of chlamydiae by nested PCR. In: K. Sachse and J. Frey (eds), PCR Detection of Microbial Pathogens: Methods and Protocols. Methods in Molecular Biology, vol. 216 (Humana Press, Totowa, NJ), 231–246

  20. Schindarow, L. and Sawow, S., 1966. Cultivation of sheep abortion virus in peritoneal tissue culture of a cold-blooded animal (Testudo graeca). Zentralblatt für Veterinärmedizin B, 13, 392–397 (in German).

  21. Thompson, P.E. and Huff, C.G., 1944. Saurian malarial parasites of the United States of Mexico. Journal of Infectious Diseases, 79, 68–79

  22. Vanrompay, D., De Meurichy, W., Ducatelle, R. and Haesebrouck, F., 1994. Pneumonia in Moorish tortoises (Testudo graeca) associated with avian serovar A Chlamydia psittaci. Veterinary Record, 135, 284–285

  23. Wardrop, S., Fowler, A., O’Callaghan, P., Giffard, P. and Timms, P., 1999. Characterization of the koala biovar of Chlamydia pneumoniae at four gene loci — ompA. VD4, ompB, 16S rRNA, groESL spacer region. Systematic and Applied Microbiology, 22, 22–27

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Correspondence to K. Sachse.

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Hotzel, H., Blahak, S., Diller, R. et al. Evidence of infection in tortoises by Chlamydia-like organisms that are genetically distinct from known Chlamydiaceae species. Vet Res Commun 29, 71–80 (2005).

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  • Chlamydiaceae
  • DNA sequencing
  • genetic relatedness
  • PCR
  • Testudo spp.
  • tortoise