From April 2002 through April 2005, southern, checkered, and bandtail puffer fish (n = 516) were harvested via a range of fishing gear from the original source locations of the PFP incidents in the northern and central IRL (Figure 1).
Within several weeks of collection, we thawed frozen puffer fish, measured standard lengths and wet weights, and removed skin, liver, stomach, intestinal tract, muscle, and gonads.
At various stages of this survey, we tested puffer fish tissues for STX bioactivity using the standard MBA, Ridascreen ELISA (R-Biopharm GmBH, Darmstadt, Germany), MIST Alert (Jellet Biotek, Dartmouth, Canada) PSP kit, and mouse neuroblastoma cytotoxicity assay (MNCA, Neuro-2A) and receptor-binding assay (RBA) [Association of Official Analytical Chemists (AOAC) 1990; Cembella et al.
After the initial 2002 saxitoxin puffer fish poisoning (SPFP) events, 11 southern puffer fish were divided into the tissue compartments (listed above), and tissue samples were extracted by boiling in 0.
Puffer fish mucus or Pyrodinium samples on filters were homogenized in 0.
All three assays (MBA, MNCA, and RBA) confirmed elevated concentrations of toxic activity in the muscle compared to the liver (5- to 20-fold) of 11 southern puffer fish (Table 2).
Skin mucus of IRL southern puffer fish held captive for 1 year was highly toxic (2,407-9,039 [micro]g STX eq/100 g) compared with that of Florida Gulf Coast southern puffer fish (6.
Toxin profiles in unconsumed puffer fish fillets (n = 4) from a 2004 PFP event were confirmed by HPLC (Figure 2A,B) and LC-MS (Figure 2C-F) to be STX (92.
We also detected TTX (quantified by MBA and confirmed by LC-MS) in IRL checkered puffer fish (n = 3) at concentrations of 1,553 [+ or -] 919 and 53,700 [+ or -] 19,212 [micro]g TTX/100 g in the muscle and liver, respectively (Figure 3A-D).
2001), but the IRL incidents are the first in which STX poisoning has been confirmed in puffer fish originating in the United States (Quilliam et al.
2000) have found both toxin groups co-occurring in puffer fish species previously thought to contain only TTX.