Eutrophication has no short-term effect on the Cymbastela stipitata holobiont

Levels of nitrogen in coastal areas have been rapidly increasing due to accumulative inputs of sewage and terrigenous sediments carrying fertilizers. Sponges have an immense filtering capacity and may be directly impacted (positively or negatively) by elevated concentrations of nitrogen. Sponges also host a wide diversity of microbes involved in nitrogen metabolism, yet little is known about the effects of nitrogen loading on these symbiotic partnerships. Manipulative experiments were undertaken to examine the potential effects of excess nitrogen (up to 240 μM) on microbial symbiosis in the abundant sponge species Cymbastela stipitata. Microbial composition and activity were examined using 454-pyrotag sequencing of DNA- and RNA-derived samples. Despite the high levels of nitrogen exposure (up to 124-fold above ambient), sponges appeared visibly unaffected at all treatment concentrations. At the phylum level, the microbial community was consistent between all sponge samples regardless of nitrogen treatment, with Cyanobacteria and Thaumarchaeota being the dominant taxa. Higher microbial diversity was observed at the operational taxonomic units (OTU) level (97% sequence similarity), with only 40% of OTUs shared between samples from all treatments. However, a single cyanobacterial OTU dominated the community of all individuals (average 73.5%) and this OTU did not vary with nitrogen treatment. The conserved microbial community in all sponges irrespective of nitrogen treatment highlights the stability of the sponge-microbe relationship and indicates that the holobiont is resistant to short pulses of nitrogen at levels mimicking sewage effluent.