I need to study the bacterial origin of tetrodotoxin from my puffer fish. When I am isolating different cultures from the puffer fish, can I directly inject the supernatants of the bacterial cultures into the mice to check the presence of toxin? Or I should purify the toxin from the culture by chromatographic techniques and then inject it into the mice? Can anybody suggest me to do my work relatively simpler?
I really do not know and can only guess. This seems to be an interesting project to look for bacterial strains producing the toxin. Injection of the supernatants may work, but you should start with a standardized or positive control method to really detect the toxin - then you may look for a cheaper (or faster, or more suitable) method to do your screening or prescreening of the toxin. Since the toxin may not be very well water soluble (but I do not really know) you may miss some strains with your approach to find the right amount in an active form in the supernatant. Could a bacterial lysate or extract be better to start with and perhaps use some enrichment step of the putative toxin in something which could solve it better than water? These are just my guesses, I am not a bacteriologist or toxicologist.
I think it depends on the toxin type it is it endotoxin or exotoxin....go according to that...
Tetrodotoxin (TTX) is a potent neurotoxin which is known to occur in numerous taxa, including newts. The origin of TTX is unknown, but production by symbiotic bacteria is suspected for some groups.Fish poisoning by consumption of members of the order Tetraodontiformes is one of the most violent intoxications from marine species. The gonads, liver, intestines, and skin of pufferfish can contain levels of tetrodotoxin sufficient to produce rapid and violent death. The flesh of many pufferfish may not usually be dangerously toxic.
chromatographic purification of sugar like compounds is really painful. so I guess you should try the direct method. if ttx is really potent that should work.
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