Bans on overtime shifts, working with contractors and other industrial action have seen South Coast line trains cancelled on Tuesday.
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Trains between Wollongong, Kiama and Bomaderry have not been running due to the industrial action organised by the Rail, Tram and Bus Union NSW.
Replacement buses were used to transport passengers until the train services resumed in the mid-afternoon.
The action includes a ban on working standalone overtime shifts, a ban on working with contractors - to prevent NSW and Sydney Trains for filling members' overtime with contractors - and a move to have drivers and guards sign on only at their home station.
It will continue until February 21.
"We have been negotiating for many months to secure a fair new enterprise agreement, but workers' basic asks around hygiene and safety are being knocked back at every opportunity," RTBU NSW secretary Alex Claassens said.
"We'd hoped that after the industrial actions taken last year, the government and transport bureaucrats had finally agreed to listen and negotiate."
Mr Claassens said the actions aimed to minimise the impact on services.
Sydney and NSW Trains employees are negotiating a new enterprise agreement to replace the one that expired last May.
The sticking points include the maintenance of current level of hygiene with directly employed cleaners, a guarantee that any changes to services will not leave them less safe, and a commitment to jobs and services should privatisation occur.
Sydney Trains chief executive Matt Longland said it was disappointing that further industrial action was underway when staffing levels were already affected by COVID-19.
"We are working hard to limit the impact for customers and school students, so it is clear this is not the time for industrial action," Mr Longland said.
"We urge the unions to focus on bargaining at the negotiation table."
Mr Longland said train passengers could expect impacts on trains until February 21 but they would receive as much information as possible to plan their journeys.
It is not yet known whether there will be any further disruption to services in the Illawarra on Wednesday.
"The nature of industrial action means we do not know the exact extent or timing of the disruption that will take place, and therefore it makes it more difficult for us to plan for our customers as services may change at short notice," a Transport for NSW spokesperson said.
"Public transport customers should visit transportnsw.info for the latest information and consider alterative transport options."
Trackwork is also affecting trains on the South Coast line.
For three nights from Tuesday, February 8, trains will not stop at Wolli Creek, but will make an extra stop at Arncliffe between 9.50pm and 1.30am.
Buses will run between Wolli Creek and Sydenham, and Wolli Creek and Arncliffe.
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