Twelve strains of filamentous bacteria morphologically identified as 'Microthrix parvicella' were isolated from industrial activated sludge wastewater treatment plants. 16S rRNA gene sequences analysis showed that these strains were all closely related to 'Candidatus Microthrix parvicella'. Six of them, however, had a 16S rRNA gene similarity of only 95.7% and 96.7% to 'Candidatus Microthrix parvicella' suggesting the presence of a new species. The name 'Candidatus Microthrix calida' is proposed for this new microorganism. The physiological properties of these six isolates supported the description of a new taxon. The 'Candidatus Microthrix calida' strains produced thin filaments (0.3-0.7 microm diameter), they did not grow on the media supporting the growth of 'Candidatus Microthrix parvicella' and could be cultivated at higher temperature (up to 36.5 degrees C). Preliminary data on substrate uptake were obtained by microautoradiography on pure culture. Two new fluorescence in situ hybridization (FISH) probes, Mpa-T1-1260 specific for 'Candidatus Microthrix calida' and Mpa-all-1410 targeting both Microthrix species, were designed. The presence of Microthrix spp. was investigated in 114 activated sludge plants. 'Microthrix parvicella' morphotype was detected in 23% of the analysed samples and FISH analysis revealed that 'Candidatus Microthrix calida' was present in 5% of them. The remaining 'M. parvicella' filaments were positive with probe Mpa-all-1410 but could not all be identified as 'Candidatus Microthrix parvicella' suggesting the presence of more hitherto undescribed biodiversity within this morphotype.