COACHES replaced trains on the Bendigo, Swan Hill, and Echuca lines for 24 hours on Friday as V/Line workers again took industrial action over working conditions.
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The union and V/Line have been in negotiations since January last year.
Friday's action follows V/Line strikes in December.
The Rail, Tram and Bus Union has planned industrial action on other regional lines, including Maryborough, on February 18.
But V/Line chief executive James Pinder said earlier this week he hoped an agreement could be reached before further strikes.
He said V/Line had a good package on offer that considered the fact it was a fast-growing railway.
"This is not about paying people any less or taking away allowances. This is all about making sure we create the V/Line that all Victorians want for the future," he said.
Union branch secretary Luba Grigorovitch said it had long campaigned for additional funding for regional services.
"The government is expecting V/Line workers to do more with less, more services less support, and our members just won't stand for it," she said.
