Holly Bowles passed away with her parents by her bedside in a Bangkok hospital on Friday, the 19-year-old's father Shaun told 7News.
Her death comes one day after Bianca Jones, also 19, died in a different Bangkok hospital, with her family close by.
The pair, from Beaumaris in Melbourne's southeast, were on holiday in Laos last week when they became ill along with a dozen other tourists in Vang Vieng.
Both girls were heavily involved in the Beaumaris Football Club where a "private get together" was planned for players and coaches on Friday night.
"It's been an incredibly tough week for the club and tonight's event is about supporting each other through this tough time," a spokesman said.
"Particularly those girls who played with Bianca and Holly, all of whom are now playing Senior footy."
The club issued a tribute after Ms Jones' death, encouraging members to support one another.
"By their deeds and involvement in the girls and women's football programs, both Holly and Bianca have become cherished and highly respected members of the Beaumaris Sharks family," it said in a statement.
"Now more than ever, the wider Sharks Community needs to lean on each other for both comfort and support."
On November 11, Ms Jones and Ms Bowles planned to stay at the Nana Backpacker Hostel in Vang Vieng and have a few drinks but decided to go to Jaidee Bar.
Ms Jones and Ms Bowles didn't leave their dorm room for 24 hours afterwards and reported feeling unwell on November 13, before being taken to separate hospitals in Bangkok.
A British woman - Simone White, 28, a lawyer from southeast London - was the fifth foreigner to die after the suspected methanol poisoning event in Laos.
It's believed all of the affected tourists, including the two Australians, consumed drinks laced with methanol.
Thai authorities have confirmed Ms Jones had died of "brain swelling due to high levels of methanol found in her system".
Meanwhile, Laotian police have detained the manager and owner of the hostel where the Australians were staying.
An officer at Vang Vieng's Tourism Police office, who refused to give his name, told The Associated Press on Friday a "number of people" had been detained in the case but that no charges have yet been filed.
The Australian government has updated its travel advice for Laos, telling residents to be alert to the potential risk of spirit-based drinks including cocktails.
Foreign Minister Penny Wong said drink-spiking and methanol poisoning were "far too common in many parts of the world".
"At this time I would say to parents, to young people, please have a conversation about risks, please inform yourselves, please let's work together to ensure this tragedy does not happen again," she said.
One New Zealand citizen has also fallen ill in Laos and could also be a victim of methanol poisoning.
