Hostname: page-component-848d4c4894-8bljj Total loading time: 0 Render date: 2024-06-22T14:18:44.908Z Has data issue: false hasContentIssue false

Selection of specific genotypes of giardia intestinalis by growth in vitro and in vivo

Published online by Cambridge University Press:  06 April 2009

R. H. Andrews
Affiliation:
Department of Microbiology and Immunology, University of Adelaide, GPO Box 498, Adelaide, South Australia 5001, Australia
N. B. Chilton
Affiliation:
Evolutionary Biology Unit, South Australian Museum, North Terrace, Adelaide, South Australia 5000, Australia
G. Mayrhofer
Affiliation:
Department of Microbiology and Immunology, University of Adelaide, GPO Box 498, Adelaide, South Australia 5001, Australia

Summary

This study examined whether allelic changes observed when clinical isolates of Giardia intestinalis made in suckling mice were adapted to in vitro growth occurred as a result of gene switching (alternate isoenzymes) or through selection of organisms with different genotypes from mixed infections. Samples were compared electrophoretically at 20 enzyme loci. Marked allelic differences were detected between the uncloned clinical isolates grown in mice and the axenic cultures established from them. Furthermore, the allelic profiles of the uncloned isolates changed during the course of in vivo or in vitro growth. In contrast, all clones produced from each isolate retained identical allelic profiles, regardless of whether they were grown in vivo or in vitro. These findings argue against gene switching as an explanation for the observed allozyme changes and support preferential selection of organisms with specific genotypes by growth conditions. The data indicate the presence of at least 2 and possibly up to 4 distinct genotypes within each clinical isolate. The genetic differences detected between clinical isolates in suckling mice were of similar magnitude to those that separate different axenic isolates of G. intestinalis into cryptic species. Conversely, the genetic differences between the isolates were limited when sampled after establishment in vitro. These findings have significant implications for research on Giardia and other medically important parasites and raise the possibility that culture may exert a similar selective bias on the genotypes isolated from infections with other parasitic protozoa.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1992

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Abaza, S. M., Sullivan, J. J. & Visvesvara, G. S. (1991). Isoenzyme profiles of four strains of Giardia lamblia and their infectivity to jirds. American Journal of Tropical Medicine and Hygiene 44, 63–.CrossRefGoogle ScholarPubMed
Aggarwal, A., Bhatia, A., Naik, S. R. & Vinayak, V. K. (1983). Variable virulence of isolates of Giardia lamblia in mice. Annals of Tropical Medicine and Hygiene 77, 163–7.Google ScholarPubMed
Andrews, R. H., Adams, M., Boreham, P. F. L., Mayrhofer, G. & Meloni, B. P. (1989). Giardia intestinalis: electrophoretic evidence for a species complex. International Journal for Parasitology 19, 183–90.CrossRefGoogle ScholarPubMed
Andrews, R. H., Handman, E., Adams, M., Baverstock, P. R. & Mitchell, G. F. (1988). Genetic characterization of Leishmania isolates at 37 enzyme loci. International Journal for Parasitology 18, 445–52.CrossRefGoogle ScholarPubMed
Andrews, R. H., Mayrhofer, G., Chilton, N. B., Boreham, P. F. L. & Grimmond, T. R. (1992). Changes in allozyme pattern of the protozoan parasite. Giardia intestinalis. International Journal for Parasitology 22, 403–6.CrossRefGoogle ScholarPubMed
Baveja, U. K., Jyoti, A. S., Kaur, M., Agarwal, D. S., Anand, B. S. & Nanda, R. (1986). Isoenzyme studies of Giardia lamblia isolated from symptomatic cases. Australian Journal of Experimental Biology and Medical Science 64, 119–26.CrossRefGoogle ScholarPubMed
Bertram, M. E., Meyer, E. A., Lile, J. D. & Morse, S. A. (1983). A comparison of isoenzymes of five axenic Giardia isolates. Journal of Parasitology 69, 793801.CrossRefGoogle ScholarPubMed
Boreham, P. F. L., Phillips, R. E. & Shepherd, R. W. (1984). The sensitivity of Giardia intestinalis to drugs in vitro. Journal of Antimicrobial Chemotherapy 14, 449–61.CrossRefGoogle ScholarPubMed
Boreham, P. F. L., Phillips, R. E. & Shepherd, R. W. (1987). Heterogeneity in the responses of clones of Giardia intestinalis to anti-giardial drugs. Transactions of the Royal Society of Tropical Medicine and Hygiene 81, 406–7.CrossRefGoogle ScholarPubMed
Boreham, P. F. L., Phillips, R. E. & Shepherd, R. W. (1988). Altered uptake of metronidazole in vitro by stocks of Giardia intestinalis with different drug sensitivities. Transactions of the Royal Society of Tropical Medicine and Hygiene 82, 104–6.CrossRefGoogle ScholarPubMed
Cedillo-Riveria, R., Enciso-Moreno, J. A., Martinez-Palomo, A. & Ortega-Pierres, G. (1989). Giardia lamblia: isoenzyme analysis of 19 axenic strains isolated from symptomatic and asymptomatic patients in Mexico. Transactions of the Royal Society of Tropical Medicine and Hygiene 83, 644–6.CrossRefGoogle Scholar
Creasy, A., Fenton, B., Walker, A., Thaithong, S., Oliveira, S., Mutambu, S. & Walliker, D. (1990). Genetic diversity of Plasmodium falciparum shows geographic variation. American Journal of Tropical Medicine and Hygiene 42, 403–13.CrossRefGoogle Scholar
Davidson, R. A. (1984). Issues in clinical parasitology: the treatment of giardiasis. American Journal of Gastroenterology 79, 256–61.Google ScholarPubMed
Deane, M. P., Souza, M. A., Pereira, N. M., Goncalves, A. M., Momen, H. & Morel, C. M. (1984). Trypanosoma cruzi: inoculation schedules and re-isolation methods select individual strains from doubly infected mice as demonstrated by schizodeme analysis. Journal of Protozoology 31, 276–80.CrossRefGoogle Scholar
Korman, S. H., Le Blancq, S. M., Spira, D. T., ElON, J. ON, J., Reifen, R. & Deckelbaum, R. J. (1986). Giardia lamblia: identification of different strains from man. Zeitschrift für Parasitenkunde 72, 173–80.CrossRefGoogle ScholarPubMed
Kruetzer, R. D., Semko, M. E., Hendricks, L. D. & Wright, N. (1983). Identification of Leishmania spp. by multiple isozyme analysis. American Journal of Tropical Medicine and Hygiene 32, 703–15.CrossRefGoogle Scholar
Levi, G. C., De Avilia, C. A., Neto, V. A. (1977). Efficacy of various drugs for treatment of giardiasis. A comparative study. American Journal of Tropical Medicine and Hygiene 26, 564–5.CrossRefGoogle ScholarPubMed
Mcintyre, P. L., Boreham, P. F. L., Phillips, R. E. & Shepherd, R. W. (1986). Chemotherapy in giardiasis: clinical responses and in vitro drug sensitivity of human isolates in axenic culture. Journal of Pediatrics 108, 1005–10.CrossRefGoogle ScholarPubMed
Mayrhofer, G., Andrews, R. H., Ey, P. L., Albert, M. J., Grimmond, T. R. & Merry, D. J. (1992). The use of suckling mice to isolate and grow Giardia from mammalian faecal specimens for genetic analysis. Parasitology 105, 255–63.CrossRefGoogle ScholarPubMed
Meloni, B. P., Lymbery, A. J. & Thompson, R. C. A. (1988). Isoenzyme electrophoresis of 30 isolates of Giardia from humans and felines. American Journal of Tropical Medicine and Hygiene 38, 6573.CrossRefGoogle ScholarPubMed
Meloni, B. P. & Thompson, R. C. A. (1987). Comparative studies on the axenic in vitro cultivation of Giardia of human and canine origin: evidence for intraspecific variation. Transactions of the Royal Society of Tropical Medicine and Hygiene 81, 637–40.CrossRefGoogle ScholarPubMed
Mirelman, D. (1987). Effect of culture conditions and bacterial associates on the zymodemes of Entamoeba histolytica. Parasitology Today 3, 3740.CrossRefGoogle ScholarPubMed
Mirelman, D., Bracha, R., Wexler, A. & Chayen, A. (1986 a). Changes in isoenzyme patterns of a cloned culture of nonpathogenic Entamoeba histolytica during axenization. Infection and Immunity 54, 827–32.CrossRefGoogle ScholarPubMed
Mirelman, D., Bracha, R., Chayen, A., Aust-Kettis, A. & Diamond, L. S. (1986 b). Entamoeba histolytica: effect of growth conditions and bacterial associates on isoenzyme patterns and virulence. Experimental Parasitology 62, 142–8.CrossRefGoogle ScholarPubMed
Morel, C. M., Chiari, E., PlessmannCamargo, E. Camargo, E., Mattei, D. M., Romanha, A. J. & Simpson, L. (1980). Strains and clones of Trypanosoma cruzi can be characterized by patterns of restriction endonuclease products of kinetoplast DNA minicircles. Proceedings of the National Academy of Sciences, USA 7, 6810–14.CrossRefGoogle Scholar
Nash, T. E. (1989). Antigenic variation in Giardia lamblia. Experimental Parasitology 68, 238–41.CrossRefGoogle ScholarPubMed
Nash, T. E. & Keister, D. (1985). Differences in excretory–secretory products and surface antigens among 19 isolates of Giardia. Journal of Infectious Diseases 156, 974–84.CrossRefGoogle Scholar
Nash, T. E., Mccutchan, T., Keister, D., Dame, S. B., Conrad, J. D. & Gillin, F. D. (1985). Restriction-endonuclease analysis of DNA from 15 Giardia isolates obtained from humans and animals. Journal of Infectious Diseases 152, 6473.CrossRefGoogle ScholarPubMed
Pacheco, R. S., Grimaldi, G. JR., Momen, H. & Morel, C. M. (1990). Population heterogeneity among clones of New World Leishmania species. Parasitology 100, 393–8.CrossRefGoogle ScholarPubMed
Proctor, E. M., ISAAC-Renton, J. L., Boyd, J., Wong, Q. & Bowie, W. R. (1989). Isoenzyme analysis of human and animal isolates of Giardia duodenalis from British Columbia, Canada. American Journal of Tropical Medicine and Hygiene 41, 411–15.CrossRefGoogle ScholarPubMed
Rendtorff, R. (1954). The experimental transmission of human intestinal protozoan parasites. II. Giardia lamblia cysts given in capsules. American Journal of Hygiene 59, 209–20.Google ScholarPubMed
Richardson, B. J., Baverstock, P. R. & Adams, M. (1986). Allozyme Electrophoresis. A Handbook for Animal Systematics and Population Studies. Sydney: Academic Press.Google Scholar
Sargeaunt, P. G. (1987). The reliability of Entamoeba histolytica zymodemes in clinical diagnosis. Parasitology Today 3, 40–3.CrossRefGoogle ScholarPubMed
Upcroft, J. A., Capon, A. G., Dharmkrong-At, A., Upcroft, P. & Boreham, P. F. L. (1987). The partial characterization of an immunodominant antigen of Giardia intestinalis. In Advances in Giardia Research (ed. Wallis, P. & Hammond, B.). Calgary: University of Calgary Press.Google Scholar
Upcroft, J. A., Boreham, P. F. L. & Upcroft, P. (1989). Geographic variation in Giardia karyotypes. International Journal for Parasitology 19, 519–27.CrossRefGoogle ScholarPubMed
Ward, D. M., Weller, R. & Bateson, M. M. (1990). 16S rRNA sequences reveal numerous uncultured microorganisms in a natural community. Nature, London 345, 63–5.CrossRefGoogle Scholar