Variability of tetrodotoxin and of its analogues in the red-spotted newt, Notophthalmus viridescens (Amphibia: Urodela: Salamandridae)

Efts and adult specimens (n=142) of the red-spotted newt Notophthalmus viridescens from various locations in Canada and USA were analyzed for the presence of tetrodotoxin (TTX) and of its analogues 6. -epitetrodotoxin and 11-oxotetrodotoxin. Considerable individual variations in toxin levels were found within and among populations from New Hampshire, New York, Pennsylvania, and Virginia ranging from non-detectable to 69μg TTX per g newt. TTX and its analogues were absent in efts and adults from various locations in the Canadian province Nova Scotia, the northernmost distribution of the newt, and in adults from Florida. Newts kept in captivity for several years and reared on toxin-free diet lost their toxicity. Bayesian and maximum likelihood phylogenetic analysis of specimens from the various populations using three phylogenetic markers (COI, ND2 and 16S RNA) revealed that populations from the northern states of the USA and Canada are genetically homogenous, whereas the newts from Florida exhibited a much higher level of genetic divergence. An exogenous source of TTX in the newts either via the food chain or by synthesis of symbiotic bacteria is suggested to explain the high variability and lack of TTX in certain populations.