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Newly defined conditions for the in vitro cultivation and cryopreservation of Dientamoeba fragilis: new techniques set to fast track molecular studies on this organism

Published online by Cambridge University Press:  08 July 2010

J. L. N. BARRATT ,
G. R. BANIK ,
J. HARKNESS ,
D. MARRIOTT ,
J. T. ELLIS  and
D. STARK
J. L. N. BARRATT*
Affiliation:
Division of Microbiology, SydPath, St Vincent's Hospital, Darlinghurst, Australia University of Technology Sydney, Department of Medical and Molecular Biosciences, Broadway, Australia
G. R. BANIK
Affiliation:
Division of Microbiology, SydPath, St Vincent's Hospital, Darlinghurst, Australia University of Technology Sydney, Department of Medical and Molecular Biosciences, Broadway, Australia
J. HARKNESS
Affiliation:
Division of Microbiology, SydPath, St Vincent's Hospital, Darlinghurst, Australia University of Technology Sydney, Department of Medical and Molecular Biosciences, Broadway, Australia
D. MARRIOTT
Affiliation:
Division of Microbiology, SydPath, St Vincent's Hospital, Darlinghurst, Australia University of Technology Sydney, Department of Medical and Molecular Biosciences, Broadway, Australia
J. T. ELLIS
Affiliation:
University of Technology Sydney, Department of Medical and Molecular Biosciences, Broadway, Australia
D. STARK
Affiliation:
Division of Microbiology, SydPath, St Vincent's Hospital, Darlinghurst, Australia University of Technology Sydney, Department of Medical and Molecular Biosciences, Broadway, Australia
*
*Corresponding author: Department of Microbiology, St Vincent's Hospital, Darlinghurst 2010, NSW, Australia. Tel: +61 2 8382 9196. Fax: +612 8382 2989. E-mail: [email protected]
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Summary

Dientamoeba fragilis is a pathogen of the human gastrointestinal tract that is a common cause of diarrhoea. A paucity of knowledge on the in vitro cultivation and cryopreservation of Dientamoeba has meant that few studies have been conducted to investigate its biology. The objective of this study was to define, for the first time, in vitro culture conditions able to support the long-term in vitro growth of Dientamoeba. Also, we aimed to define a suitable method for cryopreserving viable Dientamoeba trophozoites. A modified BD medium, TYGM-9, Loeffler's slope medium, Robinson's medium, Medium 199, Trichosel and a Tritrichomonas fetus medium were compared, using cell counts, for their ability to support the growth of D. fragilis at various temperatures and atmospheric conditions. Loeffler's slope medium supported significantly better growth compared to other media. A temperature of 42°C and a microaerophilic atmosphere were also optimum for Dientamoeba growth. To our knowledge, this is the first study to describe and compare different culture media and conditions for the growth of clinical isolates of D. fragilis. This new technology will aid the development of diagnostics for dientamoebiasis as well as facilitate large-scale sequencing projects that will fast track molecular studies on D. fragilis.

Keywords

Dientamoeba fragilisin vitroculturecultivationcryopreservationculture mediaoptimization
Type
Research Article
Information
Parasitology , Volume 137 , Issue 13 , November 2010 , pp. 1867 - 1878
Copyright
Copyright © Cambridge University Press 2010

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