Seasonal and Spatial Variations in Bacterial Communities From Tetrodotoxin-Bearing and Non-tetrodotoxin-Bearing Clams

Tetrodotoxin (TTX) is one of the most potent naturally occurring compounds and is responsible for many human intoxications worldwide.Paphies australisare endemic clams to New Zealand which contain varying concentrations of TTX. Research suggests thatP. australisaccumulate the toxin exogenously, but the source remains uncertain. The aim of this study was to identify potential bacterial TTX-producers by exploring differences in bacterial communities in two organs ofP. australis: the siphon and digestive gland. Samples from the digestive glands of a non-toxic bivalveAustrovenus stutchburyithat lives amongst toxicP. australispopulations were also analyzed. Bacterial communities were characterized using 16S ribosomal RNA gene metabarcoding inP. australissourced monthly from the Hokianga Harbor, a site known to have TTX-bearing clams, for 1 year, from ten sites with varying TTX concentrations around New Zealand, and inA. stutchburyifrom the Hokianga Harbor. Tetrodotoxin was detected inP. australisfrom sites all around New Zealand and in allP. australiscollected monthly from the Hokianga Harbor. The toxin averaged 150 mu g kg(-1)over the year of sampling in the Hokianga Harbor but no TTX was detected in theA. stutchburyisamples from the same site. Bacterial species diversity differed amongst sites (p< 0.001,F= 5.9) and the diversity in siphon samples was significantly higher than in digestive glands (p< 0.001,F= 65.8). Spirochaetaceae (4-60%) and Mycoplasmataceae (16-78%) were the most abundant families in the siphons and the digestive glands, respectively. The bacterial communities were compared between sites with the lowest TTX concentrations and the Hokianga Harbor (site with the highest TTX concentrations), and the core bacterial communities from TTX-bearing individuals were analyzed. The results from both spatial and temporal studies corroborate with previous hypotheses thatVibrioandBacilluscould be responsible for the source of TTX in bivalves. The results from this study also indicate that marine cyanobacteria, in particular picocyanobacteria (e.g.,Cyanobium,Synechococcus,Pleurocapsa, andProchlorococcus), should be investigated further as potential TTX producers.