Evidence of accumulation of tetrodotoxin (TTX) in tissues and body parts of ectoparasitic copepods via their feeding on mucus of TTX-bearing pufferfish

Adults of the ectoparasitic copepod Caligus fugu found on tetrodotoxin (TTX)-bearing pufferfish such as Takifugu alboplumbeus and Takifugu flavipterus are known to accumulate TTX in body tissues and parts other than the ovaries, oviducts, eggs, and cuticles. This study aimed to demonstrate, using immunoenzymatic staining techniques, that the TTX-free planktonic/infective copepodid stage of C. fugu could accumulate TTX in the tissues after molting into the parasitic stage (chalimus I) and then fed on mucus of host puffers. All the tissues of the planktonic copepodids were completely TTX-free, whereas chalimus I copepods accumulated TTX in parts other than the cuticles, guts, and some muscles. Chalimus IV and adult copepods retained TTX in these body parts but not in the reproductive organs, which were TTX-resistant, indicating that TTX was not vertically transmitted via eggs. Non-cellular TTX-positive contents found in the guts of some chalimi and adults indicated that the copepods potentially accumulated TTX by feeding on host mucus rather than skin tissues and blood. This study revealed that the presence or absence of TTX in some body parts differed among individuals of the parasite.