They each saw off top elite cyclists from around the world, racing live online using the Zwift software platform, on a virtual road course called Watopia.
South African rider Pasio, who was also celebrating her 35th birthday, claimed Esports' first UCI rainbow jersey by pipping Australia's Sarah Gigante in a thrilling sprint finish to the women's race.
"I'm super happy. I wasn't a huge fan of virtual racing before the lockdown, but to win the first virtual world championships, I'm super proud," Pasio said after.
"There are plenty of things to come in the Esports world and I'm proud to be part of that movement. It's my birthday so it's time to celebrate!" - Ashleigh Moolman Pasio
Pasio used a virtual "Lightweight power-up" boost for the final uphill climb to finish the 50.03 km course in 1:13:27, as Gigante took second and Cecilla Hansen of Sweden third.
Next up, in the men's race there was an upset, as experienced Zwift rider Jason Osborne took the title in 1:05:15.
The German Olympian, who has qualified to compete as a rower at next year's Olympic Games in Tokyo and took part at Rio 2016, was not one of the pre-race favourites. But with 300m left in the race he timed his 'powerup' to perfection, leaving his rivals trailing.
"It was a completely new experience for me," the 26-year-old said after.
"I've raced in Zwift before but this is a much bigger event. There was a tough field of top guys and I just tried to keep calm, keep with the pack, not waste too much (energy), and left it all for the finish."
Danish duo Anders Foldager and Nicklas Pedersen took second and third respectively.
All the cyclists used identical stationary bikes and Zwift software in their own countries to take part in the world championships, organised by cycling's governing body.
As well as relying on their own physical strength and conditioning, each rider was awarded eleven "powerups" to use during the race. The two types of powerup boost were “Aero” and “Lightweight,” and riders would not know which powerup they would receive each time they were activated.
“Aero” made cyclists more aerodynamic for 15 seconds, while "Lightweight" reduced their weight in the virtual platform by 10 percent for 15 seconds. All competitors submitted their height and weight on video 72 hours before the start of the race, for it to be locked into the system.
Entries were only accepted from riders who were already registered with the UCI and part of an accredited anti-doping testing pool.
The winners receive virtual and physical rainbow bands jerseys to identify them as full UCI world champions.
Cycling esports was approved as an official UCI cycling discipline in 2018, becoming one of the first virtual disciplines to receive official status from its sport's international federation.
"What is extraordinary about cycling Esports is its ability to remove physical barriers and bring cyclists together.
"Cycling Esports is impartial to your geographical location. Cycling Esports uses technology to create opportunities for people to come together, have fun and stay fit at the same time.”
Prior to the inaugural Cycling Esports world championships, British track pursuit Olympic champion Elinor Barker told Olympic Channel, "It's the only race in which I'll ever race the Olympic and world champion on the road (Anna van der Breggen), a multiple world champion on the track (Kirsten Wild), and also a multiple Paralympic champion (Sarah Storey), in the same place; in the same race.