Singleton beekeepers are now required to register the location of their colonies or hives after a second varroa mite biosecurity zone was established in the Hunter on Tuesday.
Agriculture Minister Dugald Sanders said on Tuesday night that the deadly mite had been discovered in honey bee hives at three more properties - in Newcastle, Seaham and Bulahdelah - following its initial detection near the Port of Newcastle last week.
The discovery at Seaham has meant that the 50 kilometre notification zone has now extended to Singleton, requiring anyone with colonies or hives, including queen bees in cages and packaged bees, to notify NSW Department of Primary Industries of their location.
"This means a new 10km eradication zone, 25km for surveillance and an extended 50km biosecurity zone have been implemented, to rapidly shut down that new incursion and stop further spread," Mr Sanders said.
An emergency order remains in place prohibiting the movement of bees in the state after the mite was found at hives near the Port of Newcastle on Wednesday, June 22.
A 10km eradication zone was set up after the mite was discovered, with all hives and colonies within that zone to be eradicated.
With the detection of the mite in three new locations on Tuesday, new eradication zones have been established around those areas.
Around each eradication zone is a 25km surveillance zones, where officials are monitoring and inspecting managed and feral honey bees to limit the extent of incursions, and a 50km biosecurity zone.
Despite the spread of the mite, Mr Saunders said the expansion of the zones was "no cause for alarm".
"[It] actually shows the surveillance system is doing its job to stay on top of where this parasite is hiding," he said.
"I would like to encourage all beekeepers, both commercial and recreational, within the new or original impacted areas to please come forward for the good of the industry.
"We know the devastating impacts varroa mite will have on our honey supplies and pollination across the state, if this threat is not stopped.
"The best path forward is to report the locations of potentially impacted hives to aid our response, so we have all the information we need to deal with this as swiftly as possible."
A total of seven infested premises have now been discovered through contact tracing, including the initial detection at sentinel hives near the Port of Newcastle.
DPI is working closely with industry and will hold a briefing with them on what the eradication process will look like. These next steps will be finalised in the coming days.
Sign up for our newsletter to stay up to date.