Folia Parasitologica 50[2] 97-104 (2003) | DOI: 10.14411/fp.2003.017

Genotypic, phenotypic, biochemical, physiological and pathogenicity-based categorisation of Acanthamoeba strains

Naveed Ahmed Khan1, Noor Khan Tareen2
1 School of Biological and Chemical Sciences, Birkbeck College, University of London, London, WC1E 7HX, England, UK
2 District Headquarters Hospital, Bahawalnagar, Pakistan

The genus Acanthamoeba includes more than 20 morphological species, but classification is problematical. Recently, the discovery of substantial interstrain differences in ribosomal DNA (rDNA) sequences has prompted questions about the relatedness of strains of the same species. In this study, therefore, we have investigated relationships between two isolates of A. polyphaga, CCAP 1501/3c and ATCC 30871, using morphological, biochemical, physiological, molecular and cytotoxicity assays. We observed that A. polyphaga ATCC 30871 exhibited up to six arms in endocyst while A. polyphaga CCAP 1501/3c exhibited a maximum of 5 arms thus indicating their position in group 2 and 3, respectively. Acanthamoeba polyphaga ATCC 30871 exhibited growth at 37°C and growth on 1M mannitol plates while A. polyphaga CCAP 1501/3c did not. In addition, both isolates exhibited differences in isoenzyme banding patterns and rDNA restriction fragment polymorphisms. More importantly, A. polyphaga ATCC 30871 produced cytotoxicity on corneal epithelial cells while A. polyphaga CCAP 1501/3c had no effects, suggesting differences in pathogenicity. Thus, all the results provide evidence for significant differences between the strains and further provided the basis for reclassification of the isolates. Implications of these results in the clinical diagnosis of pathogenic Acanthamoeba are discussed.

Keywords: Acanthamoeba, cytotoxicity, differentiation, keratitis, corneal epithelial cells polyphaga, CCAP 1501/3c and ATCC 30871, using morphological, biochemical, physiological, molecular and cytotoxicity

Received: August 21, 2002; Accepted: November 19, 2002; Published: June 1, 2003  Show citation

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Khan, N.A., & Khan Tareen, N. (2003). Genotypic, phenotypic, biochemical, physiological and pathogenicity-based categorisation of Acanthamoeba strains. Folia Parasitologica, 50(2), 97-104. doi: 10.14411/fp.2003.017
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    References

    1. ABBASZADEGAN M., GERBA C.P., ROSE J.B. 1991: Detection of Giardia cysts with a cDNA probe and applications to water samples. Appl. Environ. Microbiol. 57: 927-931 Go to original source... Go to PubMed...
    2. AURAN J.D., STARR M.B., JAKOBIEC F.A. 1987: Acanthamoeba keratitis. A review of the literature. Cornea 6: 2- 26. Go to original source... Go to PubMed...
    3. BADENOCH P.R., ADAMS M., COSTER D.J. 1994: Corneal virulence, cytopathic effect on human keratocytes and genetic characterization of Acanthamoeba. Int. J. Parasitol. 25: 229-239 Go to original source... Go to PubMed...
    4. BRACHA R., DIAMOND L.S., ACKERS J.P., BURCHARD G.D., MIRELMAN D. 1990: Differentiation of clinical isolates of Entamoeba histolytica by using specific DNA probes. J. Clin. Microbiol. 28: 680-684 Go to original source... Go to PubMed...
    5. CAZENAVE J., CHEYROU A., BLOUIN A., JOHNSON M., BEGUERET J. 1991: Use of polymerase chain reaction to detect Toxoplasma. J. Clin. Pathol. 44: 1037 Go to original source... Go to PubMed...
    6. COSTAS M., GRIFFITHS A.J. 1986: Physiological characterization of Acanthamoeba isolates. J. Protozool. 33: 304-309 Go to original source...
    7. DAGGETT P.M., LIPSCOMB D.S., THOMAS K., NERAD T.A. 1985: A molecular approach to the phylogeny of Acanthamoeba. Biosystems 18: 399-405 Go to original source... Go to PubMed...
    8. DE JONCKHEERE J.F. 1980: Growth characteristics, cytopathic effect in cell culture, and virulence in mice of 36 type strains belonging to 19 different Acanthamoeba spp. Appl. Environ. Microbiol. 39: 681-685 Go to original source... Go to PubMed...
    9. DE JONCKHEERE J.F. 1983: Isoenzyme and total protein analysis by agarose isoelectric focusing and taxonomy of the genus Acanthamoeba. J. Protozool. 30: 701-706 Go to original source...
    10. DI GREGORIO C., RIVASI F., MONGIARDO N., DE RIENZO B., VISVESVARA G.S. 1991: Acanthamoeba meningoencephalitis in an AIDS patient: first report from Europe. Arch. Pathol. Lab. Med. 116: 1363-1365
    11. FRIEDLAND L.R., RAPHAEL S.A., DEUTSCH E.S., JOHAL J., MARTYN L.J., VISVESVARA G.S., LISCHNER H. 1992: Disseminated Acanthamoeba infection in a child with symptomatic human immunodeficiency virus infection. Pediatr. Infect. Dis. J. 11: 404- 407 Go to original source... Go to PubMed...
    12. GARFINKEL L., GILADI M.I., HUBER M., GITLER C., MIRELMAN D., REVEL M., ROZENBLATT S. 1989: DNA probes specific for Entamoeba histolytica possessing pathogenic and nonpathogenic zymodemes. Infect. Immun. 57: 926-931 Go to original source... Go to PubMed...
    13. GAST R.J. 2001: Development of an Acanthamoeba-specific reverse dot-blot and the discovery of a new ribotype. J. Eukaryot. Microbiol. 48: 609-615 Go to original source... Go to PubMed...
    14. GONZALEZ M.M., GOULD E., DICKINSON G., MARTINEZ A.J., VISVESVARA G.S., CLEARY T.J., HENSLEY G.T. 1986: Acquired immunodeficiency syndrome associated with Acanthamoeba infection and other opportunistic organisms. Arch. Pathol. Lab. Med. 110: 749-751
    15. GORDON S.M., STEINBERG J.P., DUPUIS M., KOZARSKY P., NICKERSON J.F., VISVESVARA G.S. 1992: Culture isolation of Acanthamoeba species and leptomyxid amebas from patients with amebic meningoencephalitis, including two patients with AIDS. Clin. Infect. Dis. 15: 1024-1030 Go to original source... Go to PubMed...
    16. GUAY J.M., DUBOIS D., MORENCY J.M., GAGNON S., MERCIER J., LEVESQUE C.R. 1993: Detection of the pathogenic parasite Toxoplasma gondii by specific amplification of ribosomal sequences using comultiplex polymerase chain reaction. J. Clin. Microbiol. 31: 203- 207 Go to original source... Go to PubMed...
    17. JOHN D.T. 1993: Opportunistically pathogenic free-living amebae. In: J.P. Kreier and J.R. Baker (Eds.), Parasitic Protozoa. Vol. 3. Academic Press, New York, pp. 143- 246 Go to original source...
    18. KHAN N.A. 2001: Pathogenicity, morphology and differentiation of Acanthamoeba. Curr. Microbiol. 43: 391-395 Go to original source... Go to PubMed...
    19. KHAN N.A., JARROLL E.L., PAGET T.A. 2001: Acanthamoeba can be differentiated clinically by the polymerase chain reaction and simple plating assays. Curr. Microbiol. 43: 204-208 Go to original source... Go to PubMed...
    20. KHAN N.A., JARROLL E.L., PANJWANI N., CAO Z., PAGET T.A. 2000: Proteases as markers of differentiation of pathogenic and non-pathogenic Acanthamoeba. J. Clin. Microbiol. 38: 2858-2861 Go to original source... Go to PubMed...
    21. KONG H.H., CHUNG D.I. 1996: PCR and RFLP variation of conserved region of small subunit ribosomal DNA among Acanthamoeba isolates assigned to either A. castellanii or A. polyphaga. Korean J. Parasitol. 34: 127-134 Go to original source... Go to PubMed...
    22. LAXER M.A., D'NICUOLA E.M., PATEL J.R. 1992: Detection of Cryptosporidium parvum DNA in fixed, paraffin-embedded tissue by the polymerase chain reaction. Am. J. Trop. Med. Hyg. 47: 450-455 Go to original source... Go to PubMed...
    23. LAXER M.A., TIMBLIN K.B., PATEL J.R. 1991: DNA sequences for the specific detection of Cryptosporidium parvum by the polymerase chain reaction. Am. J. Trop. Med. Hyg. 45: 688-694 Go to original source... Go to PubMed...
    24. MacPHERSON J.M., GAJADHAR A.A. 1993: Sensitive and specific polymerase chain reaction detection of Toxoplasma gondii for veterinary and medical diagnosis. Can. J. Vet. Res. 57: 45-48
    25. MAHBUBANI M.H., BEJ A.K., PERLIN M., SCHAEFER F.W., JAKUBOWSKI W., ATLAS R.M. 1991: Detection of Giardia cysts by using the polymerase chain reaction and distinguishing live from dead cysts. Appl. Environ. Microbiol. 57: 3456-3461 Go to original source... Go to PubMed...
    26. MAHBUBANI M.H., BEJ A.K., PERLIN M., SCHAEFER F.W., JAKUBOWSKI W., ATLAS R.M. 1992: Differentiation of Giardia duodenalis from other Giardia spp. by using polymerase chain reaction and gene probes. J. Clin. Microbiol. 30: 74-78 Go to original source... Go to PubMed...
    27. MARCIANO-CABRAL F., PUFFENBARGER R., CABRAL G.A. 2000: The increasing importance of Acanthamoeba infections. J. Eukaryot. Microbiol. 47: 29-36 Go to original source... Go to PubMed...
    28. MATHIOPOULOS K., BOUARE M., McCONKEY G., McCUTCHAN T. 1993: PCR detection of Plasmodium species in blood and mosquitoes. In: D.H. Persing, T.F. Smith, F.C. Tenover and T.J. White (Eds.), Diagnostic Molecular Microbiology. American Society for Microbiology, Washington, D.C., USA, pp. 462-467
    29. NIEDERKORN J.Y., ALIZADEH H., LEHER H., McCULLEY J.P. 1999: The pathogenesis of Acanthamoeba keratitis. Microb. Infect. 1: 437-443 Go to original source... Go to PubMed...
    30. PUSSARD M., PONS R. 1977: Morphologie de la paroi kystique et taxonomie du genre Acanthamoeba (Protozoa, Amoebida). Protistologica 13: 557-598
    31. SPARAGANO O. 1993: Differentiation of Naegleria fowleri and other Naegleriae by polymerase chain reaction and hybridization methods. FEMS Microbiol. Lett. 110: 325- 330. Go to original source... Go to PubMed...
    32. STOTHARD D.R., SCHROEDER-DIEDRICH J.M., AWWAD M.H., GAST R.J., LEDEE D.R., RODRIGUEZZARAGOZA S., DEAN C.L., FUERST P.A., BYERS T.J. 1998: The evolutionary history of the genus Acanthamoeba and the identification of eight new rRNA gene sequence types. J. Eukaryot. Microbiol. 45: 45-54 Go to original source... Go to PubMed...
    33. ULIANA S.R.B., AFFONZO M.J.T., CAMARGO E.P., FLOETER-WINTER L.M. 1991: Leishmania: genus identification based on a specific sequence of the 18S ribosomal RNA sequence. Exp. Parasitol. 72: 157-163 Go to original source... Go to PubMed...
    34. VERHOFSTEDE C., RENIERS S., COLEBUNDERS R., WANZEELE F., PLUM J. 1993: Polymerase chain reaction in the diagnosis of Toxoplasma encephalitis, AIDS 7: 1539-1541 Go to original source... Go to PubMed...
    35. VISVESVARA G.S. 1991: Classification of Acanthamoeba. Rev. Infect. Dis. 13 (S5): 369-372 Go to original source...
    36. WATERS A.P., McCUTCHAN T.F. 1989: Rapid, sensitive diagnosis of malaria based on ribosomal RNA. Lancet i: 1343-1346
    37. WEEKERS H.H.P., DE JONCKHEERE J.F. 1997: Differences in isoenzyme patterns of axenically and monoxenically grown Acanthamoeba and Hartmannella. Antonie van Leeuwenhoek 71: 231-237 Go to PubMed...
    38. YANG Z., CAO Z., PANJWANI N. 1997: Pathogenesis of Acanthamoeba keratitis: carbohydrate-mediated hostparasite interactions. Infect. Immun. 65: 439-445 Go to original source... Go to PubMed...

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