Hostname: page-component-8448b6f56d-xtgtn Total loading time: 0 Render date: 2024-04-17T17:30:07.578Z Has data issue: false hasContentIssue false

The use of suckling mice to isolate and grow Giardia from mammalian faecal specimens for genetic analysis

Published online by Cambridge University Press:  06 April 2009

G. Mayrhofer
Affiliation:
Department of Microbiology and Immunology, University of Adelaide, GPO Box 498, Adelaide SA 5001, South Australia
R. H. Andrews
Affiliation:
Department of Microbiology and Immunology, University of Adelaide, GPO Box 498, Adelaide SA 5001, South Australia
P. L. Ey
Affiliation:
Department of Microbiology and Immunology, University of Adelaide, GPO Box 498, Adelaide SA 5001, South Australia
M. J. Albert
Affiliation:
Alice Springs Hospital, Alice Springs, and Menzies School of Health Research, Darwin, Northern Territory
T. R. Grimmond
Affiliation:
Department of Clinical Microbiology, Flinders Medical Centre, Bedford Park, South Australia
D. J. Merry
Affiliation:
Division of Clinical Microbiology, Institute of Medical and Veterinary Science, Adelaide, South Australia

Summary

A simple technique is described for preparation of Giardia cysts from faecal samples, the growth of trophozoites in suckling mice and the isolation of trophozoites for genetic analysis by allozyme electrophoresis. In total, 125 new isolates of Giardia have been collected from human and animal sources over a wide geographical area of South Australia and the Northern Territory of Australia. A number of long-established axenized isolates of G. intestinalis belonging to Groups I and II also adapted to grow in suckling mice. These findings indicate that suckling mice are permissive hosts for a variety of genetically dissimilar but morphologically similar organisms of the G. duodenalis type and that this in vivo technique may be less selective than isolation by in vitro culture. The use of suckling mice has revealed that infections can be composed of mixed genotypes and that isolation and purification techniques can be selective. Allozymic interpretation is essential to reveal the genetic complexity of such mixtures

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

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., Mayrhofer, G., Chilton, N. B., Boreham, P. F. L. & Grimmond, T. R. (1992 a). Changes in allozyme pattern of the protozoan parasite Giardia intestinalis. International Journal for Parasitology (in the Press).CrossRefGoogle ScholarPubMed
Andrews, R. H., Chilton, N. B. & Mayrhofer, G. (1992 b). Selection of specific genotypes of Giardia intestinalis by growth in vitro and in vivo. Parasitology (in the Press).CrossRefGoogle ScholarPubMed
Farthing, M. J. G., Varon, S. R. & Keusch, G. T. (1983). Mammalian bile promotes growth of Giardia lamblia in axenic culture. Transactions of the Royal Society of Tropical Medicine and Hygiene 77, 467–9.CrossRefGoogle ScholarPubMed
Feely, D. E. & Erlandsen, S. L. (1981). Isolation and purification of Giardia trophozoites from rat small intestine. Journal of Parasitology 67, 5964.CrossRefGoogle Scholar
Filice, F. P. (1952). Studies on the cytology and life history of Giardia from the laboratory rat. University of California Publications in Zoology 57, 53146.Google Scholar
Fortess, E. & Meyer, E. A. (1976). Isolation and axenic cultivation of Giardia trophozoites from the guinea pig. Journal of Parasitology 62, 689.CrossRefGoogle ScholarPubMed
Gillin, F. D. & Diamond, L. S. (1981). Entamoeba histolytica and Giardia lamblia: growth responses to reducing agents. Experimental Parasitology 51, 382–91.CrossRefGoogle ScholarPubMed
Hill, D. R., Guerrant, R. L., Pearson, R. D. & Hewlett, E. L. (1983). Giardia lamblia infection of suckling mice. Journal of Infectious Diseases 147, 217–21.CrossRefGoogle ScholarPubMed
Keister, D. B. (1983). Axenic culture of Giardia lamblia in TY1-S-33 medium supplemented with bile. Transactions of the Royal Society of Tropical Medicine and Hygiene 77, 487–8.CrossRefGoogle Scholar
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
Meyer, E. A. (1970). Isolation and axenic culture of Giardia trophozoites from the rabbit, chinchilla and cat. Experimental Parasitology 27, 179–83.CrossRefGoogle ScholarPubMed
Meyer, E. A. (1976). Giardia lamblia: isolation and axenic cultivation. Experimental Parasitology 39, 101–5.CrossRefGoogle ScholarPubMed
Phillips, R. E., Boreham, P. F. L. & Shepherd, R. W. (1984). Cryopreservation of viable Gardia intestinalis trophozoites. Transactions of the Royal Society of Tropical Medicine and Hygiene 78, 604–6.CrossRefGoogle Scholar
Richardson, B. J., Baverstock, P. R. & Adams, M. (1986). Allozyme Electrophoresis: A Handbook for Animal Systematics and Population Studies. Sydney: Academic Press.Google Scholar
Roberts-Thomson, I. C., Stevens, D. P., Mahmoud, A. A. F. & Warren, K. S. (1976). Giardiasis in the mouse: an animal model. Gastroenterology 71, 5761.CrossRefGoogle ScholarPubMed
Sauch, J. F. (1984). Purification of Giardia muris by velocity sedimentation. Applied Environmental Microbiology 48, 454–5.CrossRefGoogle ScholarPubMed
Thompson, R. C. A., Lymbery, A. J. & Meloni, B. P. (1990). Genetic variation in Giardia Kunstler, 1882: taxonomic and epidemiological significance. Protozoological Abstracts 14, 128.Google Scholar
Visvesvara, G. S. (1980). Axenic growth of Giardia lamblia in Diamond's TYS-1 medium. Transactions of the Royal Society of Tropical Medicine and Hygiene 2 1315.Google Scholar
Waight Sharma, A. & Mayrhofer, G. (1988). A comparative study of infections with rodent isolates of Giardia duodenalis in inbred strains of rats and mice in hypothymic nude rats. Parasite Immunology 10, 169–79.CrossRefGoogle Scholar
Ward, D. M., Weller, R. & Bateson, M. M. (1990). 16S RNA sequences reveal numerous uncultured microorganisms in a natural community. Nature, London 345, 63–5.CrossRefGoogle Scholar