Hostname: page-component-78c5997874-fbnjt Total loading time: 0 Render date: 2024-11-10T14:25:16.128Z Has data issue: false hasContentIssue false

Clonal cultures of Histomonas meleagridis, Tetratrichomonas gallinarum and a Blastocystis sp. established through micromanipulation

Published online by Cambridge University Press:  20 July 2006

M. HESS
Affiliation:
Clinic for Avian, Reptile and Fish Medicine, University of Veterinary Medicine, Vienna, Veterinärplatz 1, 1210 Vienna, Austria
T. KOLBE
Affiliation:
Department Biotechnology in Animal Production, Institute for Agrobiotechnology, Konrad Lorenz Straße 20, 3430 Tulln, Austria
E. GRABENSTEINER
Affiliation:
Clinic for Avian, Reptile and Fish Medicine, University of Veterinary Medicine, Vienna, Veterinärplatz 1, 1210 Vienna, Austria
H. PROSL
Affiliation:
Institute of Parasitology and Zoology, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210, Vienna, Austria

Abstract

Clonal cultures of Histomonas meleagridis, Tetratrichomonas gallinarum and a Blastocystis sp. were established for the first time. Single microbes were successfully isolated from a mixture of micro-organisms obtained from caecal contents of turkeys, using a micromanipulation approach. The cloned parasites were propagated in vitro and maintained through continuous passages multiplying to high numbers. Identification of the protists was done by morphological investigation identifying various forms of each parasite. PCR and partial sequencing of the small subunit rRNA were used to confirm clonality and to determine the relationship of the cloned parasites with known protozoan parasites. The clonal cultures established by this technique will be useful to gain more insight into the biological repertoire of the organisms. In addition, refined infection experiments in different poultry species can now be performed to elucidate the pathological pathways of the respective protozoa.

Type
Research Article
Copyright
2006 Cambridge University Press

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

Allen, E. A. A. ( 1936). Pentatrichomonas associated with certain cases of enterohepatitis or “blackhead” of poultry. Transactions of the American Microscopic Society 55, 315322.CrossRefGoogle Scholar
Arisue, N., Hashimoto, T. and Yoshikawa, H. ( 2003). Sequence heterogeneity of the small subunit ribosomal RNA genes among Blastocystis isolates. Parasitology 126, 19.CrossRefGoogle Scholar
Bayon, H. P. and Bishop, A. ( 1937). Cultivation of Histomonas meleagridis from the liver lesions of a hen. Nature, London 139, 370371.CrossRefGoogle Scholar
Bushek, D., Holley, R. A. and Reece, K. S. ( 2000). Use of micromanipulation and “feeder layers” to clone the oyster pathogen Perkinsus marinus. Journal of Eukaryotic Microbiology 47, 164166.CrossRefGoogle Scholar
Cepicka, I., Kutisova, K., Tachezy, J., Kulda, J. and Flegr, J. ( 2005). Cryptic species within the Tetratrichomonas gallinarum species complex revealed by molecular polymorphism. Veterinary Parasitology 128, 1121.CrossRefGoogle Scholar
Clark, C. G. and Diamond, L. S. ( 2002). Methods for cultivation of luminal parasitic protists of clinical importance. Clinical Microbiological Reviews 15, 329341.CrossRefGoogle Scholar
Delgado-Viscogliosi, P., Viscogliosi, E., Gerbod, D., Kulda, J., Sogin, M. L. and Edgcomb, V. P. ( 2000). Molecular phylogeny of parabasalids based on small subunit rRNA sequences, with emphasis on the Trichomonadinae subfamily. Journal of Eukaryotic Microbiology 47, 7075.CrossRefGoogle Scholar
Dwyer, D. M. ( 1970). An improved method for cultivating Histomonas meleagridis. Journal of Parasitology 56, 191192.CrossRefGoogle Scholar
Farri, T. A. ( 1978). Simple technique for preparing clone cultures of Entamoeba histolytica. Transactions of the Royal Society of Tropical Medicine and Hygiene 72, 205206.CrossRefGoogle Scholar
Friedhoff, K. T., Kuhnigk, C. and Müller, I. ( 1991). Experimental infections in chickens with Chilomastix gallinarum, Tetratrichomonas gallinarum, and Tritrichomonas eberthi. Parasitological Research 77, 329334.CrossRefGoogle Scholar
Gerbod, D., Edgcomb, V. P., Noel, C., Zenner, L., Wintjens, R., Delgado-Viscogliosi, P., Holder, M. E., Sogin, M. L. and Viscogliosi, E. ( 2001). Phylogenetic position of the trichomonad parasite of turkeys, Histomonas meleagridis (Smith) Tyzzer, inferred from small subunit rRNA sequence. Journal of Eukaryotic Microbiology 48, 498504.CrossRefGoogle Scholar
Gillin, F. D. and Diamond, L. S. ( 1978). Clonal growth of Entamoeba histolytica and other species of Entamoeba in agar. Journal of Protozoology 25, 539543.CrossRefGoogle Scholar
Grabensteiner, E. and Hess, M. ( 2005). Sensitiver und spezifischer Nachweis von Histomonas meleagridis, Tetratrichomonas gallinarum und Blastocystis spp. 68. Fachgespräch über Geflügelkrankheiten, Hannover, 12.-13.05.2005., 39-44. ISBN 3-938026-48-0.
Granger, B. L., Warwood, S. J., Benchimol, M. and De Souza, W. ( 2000). Transient invagination of flagella by Tritrichomonas foetus. Parasitology Research 86, 699709.CrossRefGoogle Scholar
Hess, M., Grabensteiner, E., Liebhart, D., Weissenböck, H. and Loupal, G. ( 2004). Diagnostic investigations on Histomonas meleagridis following a severe outbreak in a turkey flock. Fifth International Symposium on Turkey Diseases, Berlin, 16–19 June 2004, pp. 254257.
Honigberg, B. M. and Brugerolle, G. ( 1990). Structure in Trichomonads parasitic in humans. In Trichomonas Parasitic in Humans ( ed. Honigberg, B. M.) pp. 535. Springer-Verlag, New York.CrossRef
Martin, C. H. and Robertson, M. ( 1911). Further observations on the caecal parasites of fowls, with some reference to the rectal fauna of other vertebrates. Quarterly Journal of Microbiological Science 57, 5381.Google Scholar
McDougald, L. R. ( 2003). Other protozoan diseases of the intestinal tract – Histomoniasis (Blackhead). In Diseases of Poultry ( ed. Saif, Y. M., Barnes, H. J., Glisson, J. R., Fadly, A. M., McDougald, L. R. and Swayne, D. E.), pp. 10011008. Iowa State Press, Ames, Iowa.
Noel, C., Peyronnet, C., Gerbod, D., Edgcomb, V. P., Delgado-Viscogliosi, P., Sogin, M. L., Capron, M., Viscogliosi, E. and Zenner, L. ( 2003). Phylogenetic analysis of Blastocystis isolates from different hosts based on the comparison of small-subunit rRNA gene sequences. Molecular and Biochemical Parasitology 126, 119123.CrossRefGoogle Scholar
Oduola, A. M. J., Weatherly, N. F., Bowdre, J. H. and Desjardins, R. E. ( 1988). Plasmodium falciparum – cloning by single-erythrocyte micromanipulation and heterogeneity in vitro. Experimental Parasitology 66, 8695.CrossRefGoogle Scholar
Shirley, M. W. and Millard, B. J. ( 1976) Some observations on the sexual differentiation of Eimeria tenella using single sporozoite infections in chicken embryos. Parasitology 73, 337341.CrossRefGoogle Scholar
Stenzel, D. J. and Boreham, P. F. L. ( 1996). Blastocystis hominis revisited. Clinical Microbiology Reviews 9, 563584.Google Scholar
Stepkowski, S. and Klimont, S. ( 1979). Obserwacje nad hodowlg in vitro Histomonas meleagridis (Smith, 1895), Medycyna Weterynaryjna 8, 502505.Google Scholar
Tan, S. W., Singh, M., Thong, K. T., Ho, L. C., Moe, K. T., Chen, X. Q., Ng, G. C. and Yap, E. H. ( 1996). Clonal growth of Blastocystis hominis in soft agar with sodium thioglycollate. Parasitology Research 82, 737739.CrossRefGoogle Scholar
Tan, S. W. ( 2004). Blastocystis in humans and animals: new insights using modern methodologies. Veterinary Parasitology 126, 121144.CrossRefGoogle Scholar
Tyzzer, E. E. ( 1919). Developmental phases of the protozoon of “blackhead” in turkeys. Journal of Medical Research 40, 130.Google Scholar
Tyzzer, E. E. ( 1920). The flagellate character and reclassification of the parasite producing ‘blackhead” in turkeys-Histomonas (gen. nov.) meleagridis (Smith). Journal of Parasitology 6, 124131.Google Scholar
Zierdt, C. H. and Williams, R. L. ( 1974). Blastocystis hominis – axenic cultivation. Experimental Parasitology 36, 233243.CrossRefGoogle Scholar