The event was the first national Canadian convention for the fans, who call themselves "bronies" — a portmanteau of "bros" and "ponies."
Nearly 800 people from across Canada and the United States attended the event, which featured a "Ponyville Idol" contest, a pajama party and various panels featuring voice actors, writers and musicians on the show.
The bronies, who came bedecked in costumes inspired by the show, say they hoped to spread the My Little Pony message of tolerance, inclusion and friendship to non-bronies who may not understand the show's appeal.
"Seeing a guy watching a show for little girls causes a lot of people to raise a red flag," said fan Daniel Collins. "'What's going on? Why are you into a show for little girls?' And I say, 'You have to see it for yourself because I cannot describe it.'"
The brony phenomenon began shortly after the first episode of My Little Pony: Friendship is Magic aired in 2010, more than 20 years after the original My Little Pony television series.
Message boards and online community groups were formed where thousands of men revealed they were fans of the show.
A recent survey conducted by a Louisiana psychologist estimates the brony herd to be about 30,000 members strong.
Last year, 4,000 fans showed up for a brony convention in New Jersey, the Associated Press reported.
"As a man, you think to yourself: 'What am I doing with my life?'" said fan Alex Atley. "As I kept on watching it…my heart, so to speak, became lighter and softer and ever since then I cannot be happier."