The video, posted on Friday, was described as homophobic by a number of LGBT+ members of the Republican party, as well as by critics on the other side of the aisle.
Seemingly making a dig at Donald Trump for not being homophobic enough, the video includes clips of the former president where he appeared to embrace the LGBT+ community in the wake of the 2016 mass shooting at gay nightclub Pulse.
The second part of the video shifts to Mr DeSantis and his own comments and policies on drag acts and transgender rights.
But, in a bizarre move, the video appears to try to portray the Florida governor as masculine, with his image spliced with images of muscular, shirtless men and several Hollywood actors, including Brad Pitt.
Although Mr Buttgieg didn’t comment directly on the video, he managed to work in a few jabs at Mr DeSantis during an intervew with CNN’s State of the Union on Sunday.
“I’m going to leave aside the strangeness of trying to prove your manhood by putting up a video that splices images of you in between oiled-up shirtless bodybuilders and just get to the bigger issue that is on my mind whenever I see this stuff in the policy space,” Mr Buttigieg told host Dana Bash.
“Which is, again, who are you trying to help? Who are you trying to make better off? And what public policy problems do you get up in the morning thinking about how to solve?”
He added that his own department’s work on flooding and infrastructure development was the focus for him and many other politicians.
“These are the kinds of problems that most of us got into government, politics, and public service in order to work on,” Mr Buttigieg said.
“And I just don’t understand the mentality of somebody who gets up in the morning thinking that he’s going to prove his worth by competing over who can make life hardest for a hard-hit community that is already so vulnerable in America.”
While Mr DeSantis sought to attack his main rival in the GOP presidential race over his LGBT+ stance, Mr Trump was widely condemned for his own anti-LGBT+ actions during his time in the White House.
Mr Trump faced repeated accusations of homophobia after failing to acknowledge Pride month, banning US embassies from hanging rainbow flags, and announcing via Twitter that he would ban transgender people from serving in the military.
Join our commenting forum
Join thought-provoking conversations, follow other Independent readers and see their replies