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The name of the episode is a reference to the phrase "once bitten, twice shy," referring to how people tend to become more reluctant to pursue a relationship after the failure of a previous one.
In the context of the episode, the title is more of a double pun; Jay is infected with Fangpyre venom after pricking his finger on a Fangpyre's fangs, and he tries to impress Nya because he fears that she won't appreciate his natural personality.
This episode marks the beginning of Jay and Nya's relationship. It also marks the last time Nya uses her "Samurai X" identity - although she will still use her technology, it will be under her own name.
As of this episode, the entire known Walker family has been infected with Fangpyre venom at some point - previously, Jay's parents were bitten by Fangpyre in "Snakebit."
Jay's Fangpyre infection appears to progress slightly differently from his parents'; his hand begins showing a green tint and scale textures before he begins developing fangs or a hissing voice. This may be due to his infection starting with a pricked finger, as it is never shown or mentioned where Ed and Edna were bitten in "Snakebit."
The Fangpyre skeleton appears to belong to a General, possibly Fangtom's predecessor.
The four weapons Lord Garmadon wields in his battle against Sensei Wu include a Nin-Jô and normal-colored versions of the Golden Star Hammer, the Staff of Light, and the Double-Bladed Dagger.