The effects of sulphide on growth and behaviour of the thiotrophic Zoothamnium niveum symbiosis

Zoothamnium niveum (Ciliophora, Oligohymenophora) is a giant, colonial marine ciliate from sulphide-rich, shallow-water habitats, obligatorily associated with the ectosymbiotic, chemoautotrophic, sulphide-oxidizing bacterium 'Candidatus Thiobios zoothamnicoli'. The aims of this study were to characterize the natural habitat and investigate growth, reproduction, survival and maintenance of the symbiosis from Corsica, France (Mediterranean Sea) using a flow-through respirometer providing stable chemical conditions. We were able to successfully cultivate the Z. niveum symbiosis during its entire lifespan and document reproduction, whereby the optimum conditions were found to range from 3 to 33 micromol l(-1) sigmaH2S in normoxic seawater. Starting with an inoculum of 13 specimens, we found up to 173 new specimens that were asexually produced after only 11 days. Observed mean lifespan of the Z. niveum colonies was approximately 11 days and mean colony size reached 51 branches, from which rapid host division rates of up to every 4.1 hours were calculated. Comparing the ectosymbiotic population from Z. niveum colonies collected from their natural habitat with those cultivated under optimal conditions, we found significant differences in the bacterial morphology and the frequency of dividing cells on distinct host parts, which is most likely caused by behaviour of the host ciliate. Applying different sulphide concentrations we revealed that the symbiosis was not able to survive without sulphide and was harmed by high sulphide conditions. To our knowledge, this study reports the first successful cultivation of a thiotrophic ectosymbiosis.