Effect of salinity stress and nitrogen depletion on growth, morphology and toxin production of freshwater cyanobacterium Microcoleus anatoxicus Stancheva & Conklin

Cyanobacterium Microcoleus anatoxicus, isolated from a coastal stream in northern California, produces both anatoxin-a (ATX) and dihydroanatoxin-a (dhATX), responsible for dog deaths, but its environmental preferences are unknown. We tested the effect of environmentally relevant stressors, e.g., salinity enrichment and nitrogen (N) depletion, on mat formation and toxicity of M. anatoxicus during the stationary growth phase in culture. Microcoleus anatoxicus showed broad salinity tolerance and the potential to enter estuaries and produce toxins in mesohaline conditions. Maximum growth was observed in oligohaline waters with salinity of 4.6 ppt. Moderate salinity stress (up to 7.8 ppt) did not affect dhATX production significantly. In contrast, higher salinity above 9.3 ppt had a detrimental effect on cell growth and significantly suppressed dhATX production. Formation of a common polysaccharide sheath covering multiple filaments was characteristic with increased salinity and may provide protection against osmotic stress. Microcoleus anatoxicus grown for 40 days in N-depleted medium formed mats with significantly elevated dhATX and increased ATX concentrations. Phycobilisome degradation was a possible acclimation response to N-limitation, as indicated by distinctly keritomized and pale cells in these cultures. In both experiments, most of the anatoxins were extracellular, probably due to toxin leaking during the stationary growth phase.