Revealing mistake: When the scientists inject the mouse with poison, they put it in a jar. The, after they place it down you can see someone's hand swap the jar for one containing a dead mouse. You can even hear it. (01:17:58)

Arachnophobia (1990)
Plot summary
Directed by: Frank Marshall
Starring: John Goodman, Jeff Daniels, Julian Sands, Harley Jane Kozak
Dr. Ross Jennings (Jeff Daniels) and his family have moved to the small town of Canaima, California to become its resident physician. Things go wrong for them as residents begin to die and evidence points to an infestation of poisonous spiders.
Doctor James Atherton: In this first generation, the original male also produced a queen, and together they will construct a primary nest which the queen will guard. But eventually, she will create reproductive offspring of her own. And when that happens, this town is dead... and the next town... and the next town... and the next one, and so on.
Trivia: Since spiders really aren't a trainable type of animal, the filmmakers used hairdryers to blow on them, in order to get them to move correctly.
Question: Which spider attacked the doctor in the barn, killing him? Was it the general, the queen, or just one of the offspring?
Answer: I think he's referring to the Brazilian doctor they brought in to help. If that's the case, it was the main spider that hitched a ride with the body.
Answer: One of the offspring. There was a brief shot of the little one crawling into the doctor's slipper.
Wrong doctor.
The question is actually referring to the entomologist played by Julian Sands, not the town medical doctor that Jeff Daniels moves in to replace.
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Answer: It definitely wasn't one of the offspring, as you can see when it jumped out at him that it was a very large spider, likely the general since they mentioned the queen would be guarding the egg sac in the wine cellar to defend it from other spiders.
Phaneron ★