Folia Parasitologica, vol. 55 (2008), issue 2

Folia Parasitologica 55[2] 81-94 (2008) | 10.14411/fp.2008.011

New species of Cryptosporidium Tyzzer, 1907 (Apicomplexa) from amphibian host: morphology, biology and phylogeny

Miloslav Jirků1,2, Andrea Valigurová3, Břetislav Koudela1,2, Jaroslav Křížek2, David Modrý1,2, Jan Šlapeta4
1 Department of Parasitology, University of Veterinary and Pharmaceutical Sciences, Palackého 1-3, 612 42 Brno, Czech Republic
2 Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Branišovská 31, 370 05 České Budějovice, Czech Republic
3 Department of Botany and Zoology, Faculty of Science, Masaryk University, Kotlářská 2, 611 37 Brno, Czech Republic
4 Faculty of Veterinary Science, University of Sydney, McMaster Building - B14, New South Wales 2006, Australia

Cryptosporidium fragile sp. n. (Apicomplexa) is described from black-spined toads, Duttaphrynus melanostictus (Schneider) (Amphibia, Anura, Bufonidae) from the Malay Peninsula. The parasitized animals were directly imported from Malaysia and harboured C. fragile at the time of arrival. Oocysts were subspherical to elliptical with irregular contour in optical section, measuring 6.2 (5.5-7.0) × 5.5 (5.0-6.5) µm. Oocyst wall was smooth and colourless in light microscopy. The endogenous development of C. fragile in the stomach of black-spined toad was analysed in detail using light and electron microscopy. Cryptosporidian developmental stages were confined to the surface of gastric epithelial cells. In transmission experiments, C. fragile has not been infective for one fish species, four amphibian species, one species of reptile and SCID mice. Full length small subunit rRNA gene sequence was obtained. Phylogenetic reconstruction revealed distinct status of C. fragile within the clade of species with gastric localisation including Cryptosporidium muris Tyzzer, 1907, Cryptosporidium serpentis Levine, 1980 and Cryptosporidium andersoni Lindsay, Upton, Owens, Morgan, Mead et Blagburn, 2000. Described characteristics differentiate C. fragile from the currently recognized Cryptosporidium species. Our experience with the description of C. fragile has led us to revise the recommended criteria for an introduction of a new Cryptosporidium species name. C. fragile is the first species described and named from an amphibian host. Its prevalence of 83% (15/18) in black-spined toads within the 3 months after importation calls for strict quarantine measures and import regulation for lower vertebrates.

Keywords: Cryptosporidium fragile, new species, frog, Duttaphrynus melanostictus, ultrastructure, host specificity, phylogeny, quarantine, global amphibian decline

Received: November 2, 2007; Accepted: January 14, 2008; Published: June 1, 2008


References

  1. AKIYOSHI D.E., DILO J., PEARSON C., CHAPMAN S., TUMWINE J., TZIPORI S. 2003: Characterization of Cryptosporidium meleagridis of human origin passaged through different host species. Infect. Immun. 71: 1828-1832 Go to original source... Go to PubMed...
  2. ALVAREZ-PELLITERO P., QUIROGA M.I., SITJA-BOBADILLA A., REDONDO M.J., PALENZUELA O., PADROS F., VAZQUEZ S., NIETO J.M. 2004: Cryptosporidium scophthalmi n. sp. (Apicomplexa: Cryptosporidiidae) from cultured turbot Scophthalmus maximus. Light and electron microscope description and histopathological study. Dis. Aquat. Org. 62: 133-145 Go to original source... Go to PubMed...
  3. ALVAREZ-PELLITERO P., SITJA-BOBADILLA A. 2002: Cryptosporidium molnari n. sp. (Apicomplexa: Cryptosporidiidae) infecting two marine fish species, Sparus aurata L. and Dicentrarchus labrax L. Int. J. Parasitol. 32: 1007-1021 Go to original source... Go to PubMed...
  4. ARCAY L., BAEZ DE BORDES E., BRUZAL E. 1995: Criptosporidiosis experimental en la escala de vertebrados. I. - Infection experimentales II. - Estudio histopatologico. Parasitol. al Dia 19: 20-29
  5. ARCAY L., BRUZAL E. 1993: Cryptosporidium en rios de Venezuela. Encuesta epidemiologica de una poblacion humana y fauna en convivencia. Parasitol. al Dia 17: 11-18 Go to PubMed...
  6. BROWNSTEIN D.G., STRANDBERG J.D., MONTALI R.J., BUSH M., FORTNER J. 1977: Cryptosporidium in snakes with hypertrophic gastritis. Vet. Pathol. 14: 606-617 Go to PubMed...
  7. CHEN X.M., KEITHLY J.S., PAYA C.V., LARUSSO N.F. 2002: Cryptosporidiosis. N. Engl. J. Med. 346: 1723-1731 Go to original source... Go to PubMed...
  8. CRANFIELD M.R., GRACZYK T.K. 2006: Cryptosporidiosis. In: D.R. Mader (Ed.), Reptile Medicine and Surgery. Second Edition. Saunders Elsevier, St. Louis, pp. 756-762
  9. CRAWSHAW G.J., MEHREN K.G. 1987: Cryptosporidiosis in ZOO and wild animals. In: Erkrankungen der Zootiere. Verhandlungsbericht des 29. Internationalen Symposiums ueber die Erkrankungen der Zootiere, Cardiff. Akademie-Verlag, Berlin, pp. 353-362
  10. CURRENT W.L., UPTON S.J., HAYNES T.B. 1986: The life cycle of Cryptosporidium baileyi n. sp. (Apicomplexa, Cryptosporidiidae) infecting chickens. J. Protozool. 33: 289-296 Go to original source... Go to PubMed...
  11. DASZAK P., CUNNINGHAM A.A., HYATT A.D. 2003: Infectious disease and amphibian population declines. Divers. Distrib. 9: 141-150 Go to original source...
  12. EGYED Z., SRETER T., SZELL Z., VARGA I. 2003: Characterization of Cryptosporidium spp. - recent developments and future needs. Vet. Parasitol. 111: 103-114 Go to original source... Go to PubMed...
  13. FAYER R., MORGAN U., UPTON S.J. 2000: Epidemiology of Cryptosporidium: transmission, detection and identification. Int. J. Parasitol. 30: 1305-1322 Go to original source... Go to PubMed...
  14. GRACZYK T.K., CRANFIELD M.R., GEITNER M.E. 1998: Multiple Cryptosporidium serpentis oocyst isolates from captive snakes are not transmissible to amphibians. J. Parasitol. 84: 1298-1300 Go to original source... Go to PubMed...
  15. GRACZYK T.K., FAYER R., CRANFIELD M.R. 1996: Cryptosporidium parvum is not transmissible to fish, amphibians, or reptiles. J. Parasitol. 82: 748-751 Go to original source... Go to PubMed...
  16. GREEN S.L., BOULEY D.M., JOSLING C.A., FAYER R. 2003: Cryptosporidiosis associated with emaciation and proliferative gastritis in a laboratory-reared South African clawed frog (Xenopus laevis). Comp. Med. 53: 81-84 Go to PubMed...
  17. GUINDON S., GASCUEL O. 2003: A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst. Biol. 52: 696-704 Go to original source... Go to PubMed...
  18. KOUDELA B., MODRY D. 1998: New species of Cryptosporidium (Apicomplexa, Cryptosporidiidae) from lizards. Folia Parasitol. 45: 93-100 Go to PubMed...
  19. LINDSAY D.S., UPTON S.J., OWENS D.S., MORGAN U.M., MEAD J.R., BLAGBURN B.L. 2000: Cryptosporidium andersoni n. sp. (Apicomplexa: Cryptosporiidae) from cattle, Bos taurus. J. Eukaryot. Microbiol. 47: 91-95 Go to original source... Go to PubMed...
  20. MASUNO K., YANAI T., HIRATA A., YONEMARU K., SAKAI H., SATOH M., MASEGI T., NAKAI Y. 2006: Morphological and immunohistochemical features of Cryptosporidium andersoni in cattle. Vet. Pathol. 43: 202-207 Go to original source... Go to PubMed...
  21. MEAD J.R., ARROWOOD M.J., SIDWELL R.W., HEALEY M.C. 1991: Chronic Cryptosporidium parvum infections in congenitally immunodeficient SCID and nude mice. J. Infect. Dis. 163: 1297-1304 Go to original source... Go to PubMed...
  22. MORGAN-RYAN U.M., FALL A., WARD L.A., HIJJAWI N., SULAIMAN I., FAYER R., THOMPSON R.C.A., OLSON M., LALA., XIAO L. 2002: Cryptosporidium hominis n. sp. (Apicomplexa: Cryptosporidiidae) from Homo sapiens. J. Eukaryot. Microbiol. 49: 433-440 Go to original source... Go to PubMed...
  23. MORRISON D.A. 2006: Phylogenetic analyses of parasites in the new millennium. Adv. Parasitol. 63: 1-124 Go to original source... Go to PubMed...
  24. OSTROVSKA K., PAPERNA I. 1990: Cryptosporidium sp. of the starred lizard Agama stellio: ultrastructure and life cycle. Parasitol. Res. 76: 712-720 Go to original source...
  25. PAPERNA I., VILENKIN M. 1996: Cryptosporidiosis in the gourami Trichogaster leeri: description of a new species and a proposal for a new genus, Piscicryptosporidium, for species infecting fish. Dis. Aquat. Org. 27: 95-101 Go to original source...
  26. POSADA D., CRANDALL K.A. 1998: MODELTEST: testing the model of DNA substitution. Bioinformatics 14: 817-818 Go to original source... Go to PubMed...
  27. REDUKER D.W., SPEER C.A., BLIXT J.A. 1985: Ultrastructure of Cryptosporidium parvum oocysts and excysting sporozoites as revealed by high resolution scanning electron microscopy. J. Protozool. 32: 708-711 Go to original source... Go to PubMed...
  28. RYAN U., O'HARA A., XIAO L. 2004: Molecular and biological characterization of a Cryptosporidium molnari-like isolate from a guppy (Poecilia reticulata). Appl. Environ. Microbiol. 70: 3761-3765 Go to original source... Go to PubMed...
  29. RYAN U.M., XIAO L., READ C., SULAIMAN I.M., MONIS P., LALA.A., FAYER R., PAVLASEK I. 2003a: A redescription of Cryptosporidium galli Pavlasek, 1999 (Apicomplexa: Cryptosporidiidae) from birds. J. Parasitol. 89: 809-813 Go to original source... Go to PubMed...
  30. RYAN U., XIAO L., READ C., ZHOU L., LALA.A., PAVLASEK I. 2003b: Identification of novel Cryptosporidium genotypes from the Czech Republic. Appl. Environ. Microbiol. 69: 4302-4307 Go to original source... Go to PubMed...
  31. SKERRATT L.F., BERGER L., SPEARE R., CASHINS S., MCDONALD K.R., PHILLOTT A.D., HINES H.B., KENYON N. 2007: Spread of chytridiomycosis has caused the rapid global decline and extinction of frogs. Ecohealth 4: 125-134 Go to original source...
  32. SLAPETA J. 2006: Cryptosporidium species found in cattle: a proposal for a new species. Trends Parasitol. 22: 469-474 Go to original source... Go to PubMed...
  33. SLAPETA J. 2008: Centenary of the genus Cryptosporidium: from morphological to molecular species identification. In: M.G. Ortega-Pierres, S. Caccio, R. Fayer, T. Mank, H. Smith and R.C.A. Thompson (Eds.), Giardia and Cryptosporidium. CABI Publishing, Morelia, Mexico. (In press.)
  34. SPANO F., PUTIGNANI L., MCLAUCHLIN J., CASEMORE D.P., CRISANTI A. 1997: PCR-RFLP analysis of the Cryptosporidium oocyst wall protein (COWP) gene discriminates between C. wrairi and C. parvum, and between C. parvum isolates of human and animal origin. FEMS Microbiol. Lett. 150: 209- 217 Go to original source... Go to PubMed...
  35. STUART S.N., CHANSON J.S., COX N.A., YOUNG B.E., RODRIGUES A.S.L., FISCHMAN D.L., WALLER R.W. 2004: Status and trends of amphibian declines and extinctions worldwide. Science 306: 1783-1786 Go to original source... Go to PubMed...
  36. SULAIMAN I.M., LALA.A., XIAO L. 2002: Molecular phylogeny and evolutionary relationships of Cryptosporidium parasites at the actin locus. J. Parasitol. 88: 388-394 Go to original source... Go to PubMed...
  37. TAMURA K., DUDLEY J., NEI M., KUMAR S. 2007: MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) Software Version 4.0. Mol. Biol. Evol. 24: 1596-1599 Go to original source... Go to PubMed...
  38. THOMPSON R.C.A., OLSON M.E., ZHU G., ENOMOTO S., ABRAHAMSEN M.S., HIJJAWI N.S. 2005: Cryptosporidium and cryptosporidiosis. Adv. Parasitol. 59: 77-158 Go to original source... Go to PubMed...
  39. TILLEY M., UPTON S.J., FREED P.S. 1990: A comparative study of the biology of Cryptosporidium serpentis and Cryptosporidium parvum (Apicomplexa: Cryptosporidiidae). J. Zoo Wildl. Med. 21: 463-467
  40. TYZZER E.E. 1907: A sporozoan found in the peptic glands of the common mouse. Proc. Soc. Exp. Biol. Med. 5: 12-13. Go to original source...
  41. TYZZER E.E. 1912: Cryptosporidium parvum (sp. nov.), a coccidium found in the small intestine of the common mouse. Arch. Protistenkd. 26: 394-412
  42. UNI S., ISEKI M., MAEKAWA T., MORIYA K., TAKADA S. 1987: Ultrastructure of Cryptosporidium muris (strain RN 66) parasitizing the murine stomach. Parasitol. Res. 74: 123-132 Go to original source... Go to PubMed...
  43. UPTON S.J., MCALLISTER C.T., FREED P.S., BARNARD S.M. 1989: Cryptosporidium spp. in wild and captive reptiles. J. Wildl. Dis. 25: 20-30 Go to original source... Go to PubMed...
  44. VALIGUROVA A., HOFMANNOVA L., KOUDELA B., VAVRA J. 2007: An ultrastructural comparison of the attachment sites between Gregarina steini and Cryptosporidium muris. J. Eukaryot. Microbiol. 54: 495-510 Go to original source... Go to PubMed...
  45. VALIGUROVA A., JIRKU M., KOUDELA B., GELNAR M., MODRY D., SLAPETA J. 2008: Cryptosporidia: epicellular parasites embraced by the host cell membrane. Int. J. Parasitol. (In press.)
  46. VETTERLING J.M., TAKEUCHI A., MADDEN P.A. 1971: Ultrastructure of Cryptosporidium wrairi from the guinea pig. J. Protozool. 18: 248-260 Go to original source... Go to PubMed...
  47. WRIGHT M.K., WHITAKER B.R. 2001: Quarantine. In: M.K. Wright and B.R. Whitaker (Eds.), Amphibian Medicine and Captive Husbandry. Krier Publishing Company, Malabar, Florida, pp. 303-307
  48. XIAO L., ESCALANTE L., YANG C., SULAIMAN I., ESCALANTE A.A., MONTALI R.J., FAYER R., LALA.A. 1999: Phylogenetic analysis of Cryptosporidium parasites based on the smallsubunit rRNA gene locus. Appl. Environ. Microbiol. 65: 1578-1583 Go to PubMed...
  49. XIAO L., FAYER R., RYAN U., UPTON S.J. 2004a: Cryptosporidium taxonomy: recent advances and implications for public health. Clin. Microbiol. Rev. 17: 72-97 Go to original source... Go to PubMed...
  50. XIAO L., LIMOR J., MORGAN U.M., SULAIMAN I.M., THOMPSON R.C.A., LALA.A. 2000: Sequence differences in the diagnostic target region of the oocyst wall protein gene of Cryptosporidium parasites. Appl. Environ. Microbiol. 66: 5499- 5502 Go to original source... Go to PubMed...
  51. XIAO L., RYAN U.M., GRACZYK T.K., LIMOR J., LI L., KOMBERT M., JUNGE R., SULAIMAN I.M., ZHOU L., ARROWOOD M.J., KOUDELA B., MODRY D., LALA.A. 2004b: Genetic diversity of Cryptosporidium spp. in captive reptiles. Appl. Environ. Microbiol. 70: 891-899 Go to original source... Go to PubMed...