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

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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.

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References

  • Aggarwal A, Bhatia A, Naik SR, Vianyak VK (1983) Variable virulence of isolates ofGiardia lamblia in mice. Ann Trop Med Parasitol 77:163–167

    PubMed  Google Scholar 

  • Andrews RH, Adams M, Boreham PFL, Mayrhofer G, Meloni BP (1989)Giardia intestinalis: electrophoretic evidence for a species complex. Int J Parasitol 19:183–190

    PubMed  Google Scholar 

  • Bertram MA, Meyer EA, Lile JD, Morse SA (1983) A comparison of isoenzymes of five axenicGiardia isolates. J Parasitol 69:793–801

    PubMed  Google Scholar 

  • Bhatia VN, Warhurst DC (1981) Hatching and subsequent cultivation of cysts ofGiardia intestinalis in Diamond's medium. J Trop Med Hyg 84:45

    PubMed  Google Scholar 

  • 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 81:406–407

    PubMed  Google Scholar 

  • 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 83:644–646

    PubMed  Google Scholar 

  • 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 24:162–167

    Google Scholar 

  • 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 39:23–28

    PubMed  Google Scholar 

  • Diamond LS, Harlow DR, Cunnick CC (1978) A new medium for the axenic cultivation ofEntamoeba histolytica and otherEntamoeba. Trans R Soc Trop Med Hyg 72:431–432

    PubMed  Google Scholar 

  • Fortress FP, Meyer EA (1976) Isolation and axenic cultivation ofGiardia trophozoites from the guinea pig. J Parasitol 62:689

    PubMed  Google Scholar 

  • Felice FP (1952) Studies on the cytology and life history of aGiardia from the laboratory rat. Univ Calif Berkeley Publ Zool 57:53–146

    Google Scholar 

  • Gordts B, Retoré P, Cadranel S, Hemelhof W, Rahman M, Butzler JP (1984) Routine culture ofGiardia lamblia trophozoites from human duodenal aspirates. Lancet II:137–138

    Google Scholar 

  • Hautus MA, Kortbeek LM, Vetter JCM, Laarman JJ (1988) In vitro excystation and subsequent axenic growth ofGiardia lamblia. Trans R Soc Trop Med Hyg 82:858–861

    PubMed  Google Scholar 

  • 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 18:4006

    PubMed  Google Scholar 

  • Kasprzak W, Majewska AC (1985) Improvement in isolation and axenic growth ofGiardia intestinalis strains. Trans R Soc Trop Med Hyg 79:551–557

    PubMed  Google Scholar 

  • Kirk-Mason KE, Turner MJ, Chakraborty PR (1988) Cloning and sequence of β-tubulin cDNA fromGiardia lamblia. Nucleic Acids Res 16:2733

    PubMed  Google Scholar 

  • 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 72:173–180

    Google Scholar 

  • 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 85:67–69

    PubMed  Google Scholar 

  • 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 108:1005–1010

    PubMed  Google Scholar 

  • McManus DP, Smith JD (1979) Isoelectric focusing of some enzymes fromEchinococcus granulosus (horse and sheep strains) andE. multilocularis. Trans R Soc Trop Med Hyg 73:259–265

    PubMed  Google Scholar 

  • 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 81:637–640

    PubMed  Google Scholar 

  • Meloni BP, Lymbery AJ, Thompson RCA (1988) Isozyme electrophoresis of 30 isolates ofGiardia from humans and felines. Am J Trop Med Hyg 38:65–73

    PubMed  Google Scholar 

  • 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 40:629–637

    PubMed  Google Scholar 

  • Meyer EA (1976)Giardia lamblia: isolation and axenic cultivation. Exp Parasitol 39:101–105

    PubMed  Google Scholar 

  • Nash TE, Keister DB (1985) Differences in excretory-secretory products and surface antigens among 19 isolates ofGiardia. J Infect Dis 152:1166–1171

    PubMed  Google Scholar 

  • 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 152:64–73

    PubMed  Google Scholar 

  • 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 109:2323–2335

    PubMed  Google Scholar 

  • 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 41:411–415

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger FS, Nicklen S, Coulsen AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    PubMed  Google Scholar 

  • Shaw CR, Prasad R (1970) Starch gel electrophoresis of enzymes — a compilation of recipes. Biochem Genet 4:297–320

    PubMed  Google Scholar 

  • Smith PD, Gillin FD, Spira WM, Nash TE (1982a) Chronic giardiasis: studies on drug sensitivity, toxin production and host immune response. Gastroenterology 83:797–803

    PubMed  Google Scholar 

  • Smith PD, Gillin FD, Kaysal NA, Nash TE (1982b) Antigenic analysis ofGiardia lamblia from Afghanistan, Puerto Rico and Oregon. Infect Immun 36:714–719

    PubMed  Google Scholar 

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    PubMed  Google Scholar 

  • Thompson RCA, Meloni BP, Lymbery AJ (1988) Humans and cats have genetically identical forms ofGiardia: evidence of a zoonotic relationship. Med J Austr 145:207–209

    Google Scholar 

  • Thompson RCA, Lymbery AJ, Meloni BP (1990) Genetic variation inGiardia Kunstler, 1882: taxonomic and epidemiological significance. Protozool Abstr 14:1–28

    Google Scholar 

  • 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 87:2414–2418

    PubMed  Google Scholar 

  • 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 34:53–62

    PubMed  Google Scholar 

  • 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 18:6531–6535

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Laboratory for Parasitology and Mycology, National Institute of Public Health and Environmental Protection, P.O. Box 1, 3720 BA, Bilthoven, The Netherlands

    W. L. Homan, F. H. J. van Enckevort, L. Limper & F. van Knapen

  2. Department of Molecular Cell Biology, University of Limburg, Maastricht, The Netherlands

    G. J. J. M. van Eys

  3. Royal Tropical Institute, Amsterdam, The Netherlands

    G. J. Schoone

  4. Department of Biology and Medical Parasitology, Academy of Medicine, Poznan, Poland

    W. Kasprzak & A. C. Majewska

Authors
  1. W. L. Homan
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  2. F. H. J. van Enckevort
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  3. L. Limper
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  4. G. J. J. M. van Eys
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  5. G. J. Schoone
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  6. W. Kasprzak
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  7. A. C. Majewska
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  8. F. van Knapen
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Homan, W.L., van Enckevort, F.H.J., Limper, L. et al. Comparison ofGiardia isolates from different laboratories by isoenzyme analysis and recombinant DNA probes. Parasitol Res 78, 316–323 (1992). https://doi.org/10.1007/BF00937090

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  • DOI: https://doi.org/10.1007/BF00937090

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