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Applied and Environmental Microbiology, November 2003, p. 6533-6540, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6533-6540.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Gamma-Proteobacteria Aquicella lusitana gen. nov., sp. nov., and Aquicella siphonis sp. nov. Infect Protozoa and Require Activated Charcoal for Growth in Laboratory Media

Paula Santos,¹ Isabel Pinhal,¹ Fred A. Rainey,² Nuno Empadinhas,³ Joana Costa,³ Barry Fields,⁴ Robert Benson,⁴ António Veríssimo,¹ and Milton S. da Costa^3*

Departamento de Zoologia and Centro de Neurociências, Universidade de Coimbra, 3004-517 Coimbra,¹ Departamento de Bioquímica and Centro de Neurociências, Universidade de Coimbra, 3001-401 Coimbra, Portugal,³ Department of Biological Sciences, Louisiana State University, Baton Rouge, Louisiana 70803,² Respiratory Diseases Branch, Centers for Disease Control and Prevention, Atlanta, Georgia 30333⁴

Received 23 May 2003/ Accepted 15 August 2003

Several isolates, belonging to two new species of the same novel genus of gamma-proteobacteria, were recovered from drilled well (borehole) and spa water at São Gemil in central Portugal. These organisms are phylogenetically most closely related to the strictly intracellular uncultured species of the genus Rickettsiella, which cause disease in arthropods, and to the facultatively intracellular species of the genus Legionella, some of which cause Legionnaires' disease and Pontiac fever. The São Gemil strains grew only on media containing charcoal, as is also true of the species of the genus Legionella. Unlike the vast majority of Legionella isolates, the new isolates did not require L-cysteine or ferric pyrophosphate for growth but like the legionellae had an absolute requirement for -ketoglutarate. Strains SGT-39^T and SGT-56 grew consistently between 30 and 43°C, while strains SGT-108^T and SGT-109 grew between 30 and 40°C. The pH ranges for growth of these organisms were surprisingly narrow: strains SGT-39^T and SGT-56 grew between pH 6.3 and 7.3, while strains SGT-108^T and SGT-109 grew between pH 6.3 and 7.0. Both organisms proliferated in the amoeba Hartmannella vermiformis but did not grow in U937 human cells. Based on 16S rRNA gene sequence analysis and physiological, biochemical, and chemical analysis we describe two new species of one novel genus; one species is represented by strain SGT-39^T, for which we propose the name Aquicella lusitana, while strain SGT-108^T represents a second species of the same genus, for which we propose the name Aquicella siphonis.

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* Corresponding author. Mailing address: Departamento de Bioquímica and Centro de Neurociências, Universidade de Coimbra, 3001-401 Coimbra, Portugal. Phone: 351-239824024. Fax: 351-239826798. E-mail: milton{at}ci.uc.pt.

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Applied and Environmental Microbiology, November 2003, p. 6533-6540, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6533-6540.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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