Parasitology Research

, Volume 78, Issue 4, pp 316–323

Comparison ofGiardia isolates from different laboratories by isoenzyme analysis and recombinant DNA probes

  • W. L. Homan
  • F. H. J. van Enckevort
  • L. Limper
  • G. J. J. M. van Eys
  • G. J. Schoone
  • W. Kasprzak
  • A. C. Majewska
  • F. van Knapen
Original Investigations

Abstract

A total of 13 newGiardia isolates were established in axenic culture. All of the new isolates were obtained by excystation ofGiardia cysts from the feces of patients in Dutch hospitals. These isolates were subjected to isoenzyme and DNA analysis together with isolates from Poland, Belgium, and various other parts of the world. Isoenzyme analysis revealed that nearly all of the newly established isolates exhibited unique zymodemes. Isolates obtained from individuals from Belgium and Poland, on the other hand, displayed single zymodemes. Genomic DNA libraries were constructed from isolates belonging to the latter two zymodemes; specific and common recombinant DNA clones were selected from these libraries. Differential screening revealed that the two isolates had only 80% of the clones in common. Restriction-fragment-length polymorphism analysis using three different probes together with two synthetic probes that are complementary toGiardia structural protein genes led to the separation of all isolates into two major groups; within these groups, a further division could be made by application of other techniques or probes. The results of DNA analysis and zymodeme classification were in general agreement; in the present report they are compared with the data in the literature and discussed.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Aggarwal A, Bhatia A, Naik SR, Vianyak VK (1983) Variable virulence of isolates ofGiardia lamblia in mice. Ann Trop Med Parasitol 77:163–167PubMedGoogle Scholar
  2. Andrews RH, Adams M, Boreham PFL, Mayrhofer G, Meloni BP (1989)Giardia intestinalis: electrophoretic evidence for a species complex. Int J Parasitol 19:183–190PubMedGoogle Scholar
  3. Bertram MA, Meyer EA, Lile JD, Morse SA (1983) A comparison of isoenzymes of five axenicGiardia isolates. J Parasitol 69:793–801PubMedGoogle Scholar
  4. Bhatia VN, Warhurst DC (1981) Hatching and subsequent cultivation of cysts ofGiardia intestinalis in Diamond's medium. J Trop Med Hyg 84:45PubMedGoogle Scholar
  5. Boreham PFL, Phillips RE, Shepard RW (1987) Heterogeneity in the responses of clones ofGiardia intestinalis to anti-giardial drugs. Trans R Soc Trop Med Hyg 81:406–407PubMedGoogle Scholar
  6. Cedillo-Rivera R, Enciso-Moreno JA, Martinez-Palomo A, Ortega-Pierres G (1989)Giardia lamblia: isoenzyme analysis of 19 axenic strains isolated from symptomatic and asymptomatic patients in Mexico. Trans R Soc Trop Med Hyg 83:644–646PubMedGoogle Scholar
  7. De Jonckheere JF, Gordts B, Kasprzak W, Majewska AC, Michels PAM (1989) Cloning of a 1.8 kb repeated sequence for the identification and comparison ofGiardia intestinalis isolates. Eur J Protistol 24:162–167Google Scholar
  8. De Jonckheere JF, Majewska AC, Kasprzak W (1990)Giardia isolates from primates and rodents display the same molecular polymorphism as human isolates. Mol Biochem Parasitol 39:23–28PubMedGoogle Scholar
  9. Diamond LS, Harlow DR, Cunnick CC (1978) A new medium for the axenic cultivation ofEntamoeba histolytica and otherEntamoeba. Trans R Soc Trop Med Hyg 72:431–432PubMedGoogle Scholar
  10. Fortress FP, Meyer EA (1976) Isolation and axenic cultivation ofGiardia trophozoites from the guinea pig. J Parasitol 62:689PubMedGoogle Scholar
  11. Felice FP (1952) Studies on the cytology and life history of aGiardia from the laboratory rat. Univ Calif Berkeley Publ Zool 57:53–146Google Scholar
  12. Gordts B, Retoré P, Cadranel S, Hemelhof W, Rahman M, Butzler JP (1984) Routine culture ofGiardia lamblia trophozoites from human duodenal aspirates. Lancet II:137–138Google Scholar
  13. Hautus MA, Kortbeek LM, Vetter JCM, Laarman JJ (1988) In vitro excystation and subsequent axenic growth ofGiardia lamblia. Trans R Soc Trop Med Hyg 82:858–861PubMedGoogle Scholar
  14. Healey A, Mitchell R, Upcroft JA, Boreham PFL, Upcroft P (1990) Complete nucleotide sequence of the ribosomal RNA tandem repeat unit fromGiardia intestinalis. Nucleic Acids Res 18:4006PubMedGoogle Scholar
  15. Kasprzak W, Majewska AC (1985) Improvement in isolation and axenic growth ofGiardia intestinalis strains. Trans R Soc Trop Med Hyg 79:551–557PubMedGoogle Scholar
  16. Kirk-Mason KE, Turner MJ, Chakraborty PR (1988) Cloning and sequence of β-tubulin cDNA fromGiardia lamblia. Nucleic Acids Res 16:2733PubMedGoogle Scholar
  17. Korman SH, Le Blancq SM, Spira DT, El On J, Reifen RM, Deckelbaum RJ (1986)Giardia lamblia: identification of different strains from man. Parasitol Res 72:173–180Google Scholar
  18. Majewska AC, Kasprzak W, De Jonckheere JF, Kczmarek E (1991) Heterogeneity in the sensitivity of stocks and clones ofGiardia to metronidazole and ornodazole. Trans R Soc Trop Med Hyg 85:67–69PubMedGoogle Scholar
  19. McIntyre P, Boreham PFL, Phillips RE, Shepherd RW (1986) Chemotherapy in giardiasis: clinical responses and in vitro drug sensitivity of human isolates in axenic cultures. J Pediatr 108:1005–1010PubMedGoogle Scholar
  20. McManus DP, Smith JD (1979) Isoelectric focusing of some enzymes fromEchinococcus granulosus (horse and sheep strains) andE. multilocularis. Trans R Soc Trop Med Hyg 73:259–265PubMedGoogle Scholar
  21. Meloni BP, Thompson RCA (1987) Comparative studies on the axenic in vitro cultivation ofGiardia of human and canine origin: evidence for intraspecific variation. Trans R Soc Trop Med Hyg 81:637–640PubMedGoogle Scholar
  22. Meloni BP, Lymbery AJ, Thompson RCA (1988) Isozyme electrophoresis of 30 isolates ofGiardia from humans and felines. Am J Trop Med Hyg 38:65–73PubMedGoogle Scholar
  23. Meloni BP, Lymbery AJ, Thompson RCA (1989) Characterization ofGiardia isolates using a non-radiolabeled DNA probe, and correlation with the results of isoenzyme analysis. Am J Trop Med Hyg 40:629–637PubMedGoogle Scholar
  24. Meyer EA (1976)Giardia lamblia: isolation and axenic cultivation. Exp Parasitol 39:101–105PubMedGoogle Scholar
  25. Nash TE, Keister DB (1985) Differences in excretory-secretory products and surface antigens among 19 isolates ofGiardia. J Infect Dis 152:1166–1171PubMedGoogle Scholar
  26. Nash TE, McCutchen T, Keister D, Dame JB, Conrad JD, Gillin FD (1985) Restriction-endonuclease analysis of DNA from 15Giardia isolates obtained from humans and animals. J Infect Dis 152:64–73PubMedGoogle Scholar
  27. Peattie DA, Alonso RA, Hein A, Caulfield JP (1989) Ultrastructural localization of giardins to the edges of disk microribbons ofGiardia lamblia and the nucleotide and deduced protein sequence of alpha-giardin. J Cell Biol 109:2323–2335PubMedGoogle Scholar
  28. Proctor EM, Isaac-Renton JL, Boyd J, Wong Q, Bowie WR (1989) Isoenzyme analysis of human and animal isolates ofGiardia duodenalis from British Columbia, Canada. Am Trop Med Hyg 41:411–415Google Scholar
  29. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
  30. Sanger FS, Nicklen S, Coulsen AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467PubMedGoogle Scholar
  31. Shaw CR, Prasad R (1970) Starch gel electrophoresis of enzymes — a compilation of recipes. Biochem Genet 4:297–320PubMedGoogle Scholar
  32. Smith PD, Gillin FD, Spira WM, Nash TE (1982a) Chronic giardiasis: studies on drug sensitivity, toxin production and host immune response. Gastroenterology 83:797–803PubMedGoogle Scholar
  33. Smith PD, Gillin FD, Kaysal NA, Nash TE (1982b) Antigenic analysis ofGiardia lamblia from Afghanistan, Puerto Rico and Oregon. Infect Immun 36:714–719PubMedGoogle Scholar
  34. Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517PubMedGoogle Scholar
  35. Thompson RCA, Meloni BP, Lymbery AJ (1988) Humans and cats have genetically identical forms ofGiardia: evidence of a zoonotic relationship. Med J Austr 145:207–209Google Scholar
  36. Thompson RCA, Lymbery AJ, Meloni BP (1990) Genetic variation inGiardia Kunstler, 1882: taxonomic and epidemiological significance. Protozool Abstr 14:1–28Google Scholar
  37. Tibayrenc M, Kjellberg F, Ayala FJ (1990) A clonal theory of parasitic protozoa: the population structures ofEntamoeba, Giardia, Leishmania, Naegleria, Plasmodium, Trichomonas, andTrypanosoma and their medical and taxonomical consequences. Proc Natl Acad Sci USA 87:2414–2418PubMedGoogle Scholar
  38. Van Eys GJJM, Schoone GJ, Ligthart GS, Alvar J, Evans DA, Terpstra WJ (1989) Identification of “old world”Leishmania by recombinant DNA probes. Mol Biochem Parasitol 34:53–62PubMedGoogle Scholar
  39. Williams JGK, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • W. L. Homan
    • 1
  • F. H. J. van Enckevort
    • 1
  • L. Limper
    • 1
  • G. J. J. M. van Eys
    • 2
  • G. J. Schoone
    • 3
  • W. Kasprzak
    • 4
  • A. C. Majewska
    • 4
  • F. van Knapen
    • 1
  1. 1.Laboratory for Parasitology and MycologyNational Institute of Public Health and Environmental ProtectionBilthovenThe Netherlands
  2. 2.Department of Molecular Cell BiologyUniversity of LimburgMaastrichtThe Netherlands
  3. 3.Royal Tropical InstituteAmsterdamThe Netherlands
  4. 4.Department of Biology and Medical ParasitologyAcademy of MedicinePoznanPoland

Personalised recommendations