Carole Baskin's Dancing with the Stars debut featured a commercial about the disappearance of her late husband.
The Justice for Don Lewis advert saw the millionaire's family and their attorney, John M. Phillips, reach out to the public for answers about his disappearance in 1997, urging anyone with information to message 646-450-6530 or 1-800-litigate.
In the ad, Gale, one of the animal sanctuary owner's daughters, said: "We are a real family and to us, he was daddy."
The family is currently offering $100,000 for information about his whereabouts.
Donna, another of his daughters, explained in the ad: "I'm Donna. I'm Don Lewis' oldest daughter. We need to know what happened to our father."
The story of Baskin, 59, and Lewis came to worldwide attention through Netflix's hit documentary series Tiger King, which was released earlier this year.
Baskin - who made her Dancing with the Stars debut on Monday night (14.09.20) - has been the subject of conspiracy theories relating to the disappearance of her husband in recent months.
And in the commercial, the family's attorney referenced the speculation.
He said: "Don Lewis mysteriously disappeared in 1997. His family deserves answers, they deserve justice. Do you know who did this or if Carole Baskin was involved?"
Lewis disappeared in August 1997, when his van was found abandoned at an airport.
It's been suggested he was planning to fly to Costa Rica, but police haven't found any evidence that he ever left the US.
In April, Hillsborough County Sheriff Chad Chronister confirmed Baskin is not a suspect in his disappearance.
He said: "We don't have any evidence to even call her a person of interest."
Meanwhile, Baskin has also hit out at the way she was portrayed in Tiger King.
She explained: "I am not the money-grubbing, gold-digging, murderous person that they portrayed.
"I am the type of person who will come after any person who is abusing animals. I am relentless. I just won't stop until I find some legal way to make it stop.
"From that perspective, I think they got me right. Me as a person, that was a total assassination of my character for nothing other than whatever money they could get for selling that to Netflix."