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Kimmel was nearing the end of his speech on Cecil's death when he was overcome with emotion and could not continue.
"If you want to make this into a positive, you can uh," Kimmel said, before pausing to hold back tears. He struggled to find words and apologized before finishing his plea.
The talk show host is asking viewers to make a donation to the Wildlife Conservation Research Unit, a part of the University of Oxford Department of Zoology that had been keeping track of Cecil. Authorities in Zimbabwe identified Minnesota dentist Walter Palmer as the man facing charges for shooting and killing the lion.
The hunter issued an apology Tuesday after his kill attracted international attention. He said he wasn't aware of doing anything illegal.
"I deeply regret that my pursuit of an activity I love and practice responsibly and legally resulted in the taking of this lion," it read in part.
Like many on social media, Kimmel did not condone his actions.
"The big question is, why are you shooting a lion in the first place?" Kimmel asks early on in the video. "I'm honestly curious to know why a human being would feel compelled to do that. How is that fun?"
Kimmel said he's not against hunting if it is for a more useful cause, such as for food or to control animal populations. He hopes to bring attention to groups that protect wildlife, like the Wildlife Conservation Research Unit.
"I think it's important to have something good come out of this disgusting tragedy," he said.
Amid the increased attention, the WildCRU website was down Wednesday morning.