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Microbiology 148 (2002), 727-733
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Microbiology (2002), 148, 727-733.
© 2002 Society for General Microbiology

Research Paper

Giardia intestinalis, a eukaryote without hydrogenosomes, produces hydrogen

David Lloyd1, James R. Ralphs1 and Janine C. Harris1

Microbiology (BIOSI 1, Main Building), Cardiff University, PO Box 915, Cardiff CF10 3TL, Wales, UK1

Author for correspondence: David Lloyd. Tel: +44 29 2087 4772. Fax: +44 29 2087 4305. e-mail: LloydD{at}cardiff.ac.uk

The microaerophilic flagellated protist Giardia intestinalis, the commonest protozoal agent of intestinal infections worldwide, is of uncertain phylogeny, but is usually regarded as the earliest branching of the eukaryotic clades. Under strictly anaerobic conditions, a mass spectrometric investigation of gas production indicated a low level of generation of dihydrogen (2 nmol min-1 per 107 organisms), about 10-fold lower than that in Trichomonas vaginalis under similar conditions. Hydrogen evolution was O2 sensitive, and inhibited by 100 µM metronidazole. Fluorescent labelling of G. intestinalis cells using monoclonal antibodies to typical hydrogenosomal enzymes from T. vaginalis (malate enzyme, and succinyl-CoA synthetase {alpha} and ß subunits), and to the large-granule fraction (hydrogenosome-enriched, also from T. vaginalis) gave no discrete localization of epitopes. Cell-free extracts prepared under anaerobic conditions showed the presence of a CO-sensitive hydrogenase activity. This first report of hydrogen production in a eukaryote with no recognizable hydrogenosomes raises further questions about the early branching status of G. intestinalis; the physiological characterization of its hydrogenase, and its recently elucidated gene sequence, will aid further phylogenetic investigations.

Keywords: early branching eukaryotes, diplomonads, membrane inlet mass spectrometry, hydrogenase




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