Enchanted Forest staff were stuck in tailbacks on the A9 near Dunkeld amid ongoing disputes over the roadworks.
Earlier this week MSP John Swinney called for the roadworks to be halted after reports of schoolchildren not getting home until 6.30pm.
Organisers of the Pitlochry attraction also hit out after customers and staff were left stuck in lengthy delays on Monday.
Despite significant improvements on Tuesday, traffic issues on the stretch of the A9 have returned.
Motorists faced delays of nearly 40 minutes on Thursday according to Traffic Scotland.
A spokesperson for The Enchanted Forest said: “We’ve been hearing reports of considerably longer delays than were experienced by our visitors on Tuesday and Wednesday this week.
“We understand that BEAR Scotland has confirmed the delays have been caused by a vehicle break-down which has since been cleared.
“One of our buses has, again, been delayed on route to Pitlochry which has impacted our departures, as have some of our visitors.
“The delays and the impact on the visitor experience are very regrettable, but sadly outwith our control.”
Breakdown causes tailbacks on the A9
The works – carried out by SGN – are due to run until March and involve the installation of a new gas pipeline.
SGN explained that a breakdown close to the works caused the delays.
A spokesperson for SGN said: “We’d like to apologise for the disruption caused to motorists and residents which occurred initially when phase two of our project began on Monday.
“We’re pleased to see that traffic delays since then have been minimal and the work is continuing as planned.
“We’re aware there was a breakdown in the A9 close to our work area a short while ago, which is having a significant impact on motorists and causing delays.
“The broken down mini-bus has now been cleared from the road, and our specialist traffic management team is working to help clear the congestion.”
Rolling temporary traffic lights are set to be in place on the route for several months, except over the festive period.
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