Method for quantifying a cholinergic neurotoxin in a sample

1. A method for determining the concentration of botulinum neurotoxin sample comprising the following steps:

(a) administering the botulinum neurotoxin sample to at least one Zebra fish by intraperitoneal injection;

(b) detecting the neuromuscular activity of the botulinum neurotoxin sample,

wherein the neuromuscular activity is slowing of mobility, reduction in motility, immobility, paralysis, or a combination thereof;

(c) comparing the neuromuscular activity determined in step (b) to the neuromuscular activity of botulinum neurotoxin of known concentration; and

(d) determining the concentration of the botulinum neurotoxin sample.

2. The method of claim 1 , wherein the botulinum neurotoxin is a protein, a polypeptide, a peptide, a fusion protein, a truncated protein, a chimeric protein, a mutated protein, or a recombinant protein.

3. The method of claim 1 , wherein the botulinum neurotoxin administered in step (a) is combined with a fatty acid.

4. The method of claim 1 , wherein the Zebra fish is a fish with a wild phenotype, a mutant phenotype, or a transgenic fish.

5. The method of claim 1 , wherein one or more Zebra fish are present in a microtitration well of an automatic device.

6. The method of claim 1 , wherein step (b) is carried out with an image analyzer.

7. The method of claim 1 , wherein the neuromuscular activity is a muscular paralysis.

8. The method of claim 1 , wherein the botulinum neurotoxin is the botulinum neurotoxin of type A, type B, type C, type D, type E, type F, or type G.

9. A method for determining the concentration of botulinum neurotoxin sample comprising:

(a) administering the botulinum neurotoxin sample to at least one Zebra fish by intraperitoneal injection;

(b) detecting the biological activity of the botulinum neurotoxin sample, wherein the biological activity is slowing of mobility, reduction in motility, immobility, difference in the behavior of the fish, or a combination thereof;

(c) comparing the biological activity determined in step (b) to the biological activity of botulinum neurotoxin of known concentration; and

(d) determining the concentration of the botulinum neurotoxin sample.