"We just have so many of them -- we have an overabundance," said Rebecca Katz, of San Francisco Animal Care and Control.
The animals took off Tuesday morning from San Francisco International Airport.
They traveled in style in the main cabin of a Virgin America jet.
More than a dozen Chihuahuas were flown from San Francisco to New York City aboard Virgin America last year in the first "Operation Chihuahua" airlift. Organizers of the adoption program say the dogs quickly found new homes here.
Experts say pop culture is to blame for the overpopulation of the dogs in California, with fans imitating Chihuahua-toting celebrities like Paris Hilton.
The tiny dogs have also been sent to other states.