Mr Eavis hopes the 3.5 metre (12 feet) tall steel fence will be enough to keep out the tens of thousands of gatecrashers who have turned up in previous years - but who could now put the festival's future in jeopardy.
He has appealed to ticketless fans not to turn up again, telling them they will not be able to climb over, break down or tunnel under this year's fence.
All 100,000 tickets for this year's event have been sold - but if more people turn up, the local council could refuse permission for the festival to go ahead in future.
"For years, a lot of people have been getting into Glastonbury without tickets - over or under the fence, forgery, scams, whatever," Mr Eavis said.
"This year things have to change for good.
"We are appealing to people's consciences - please don't come without a ticket.
"The local council have made it clear that if anybody breaks through the fence, there will be no more Glastonburys."
There would be 17 hours of "excellent" coverage on BBC Two, Mr Eavis added, urging people without tickets to watch it at home instead.
Mr Eavis has held the festival on his farmland near the village of Pilton, Somerset, for more than 30 years.
He has confirmed that Rod Stewart, Isaac Hayes and Pink Floyd will be appearing this year, but the full line-up will not be announced until just nine days before the event.
Stereophonics, Coldplay and The Charlatans are among the other bands rumoured to be appearing.
Security
Some local residents have voiced concerns about what could happen if gatecrashers are turned away.
"Very little attention has been given to what goes on outside," one Pilton resident, Anne Goode, said.
"There could well be many thousands of frustrated and angry people milling around these lanes."
Police said 100,000 ticketless fans got into the three-day event in 2000 - but Mr Eavis has put that figure at 30,000.
There was no festival in 2001, and Mendip District Council made it clear that there had to be stringent security if Mr Eavis was to be granted a licence in 2002.
Fears were heightened after nine music fans were crushed to death at the Roskilde music festival in Denmark in 2000.
'Shocked'
Former Clash singer Joe Strummer lent his voice to the appeal to keep ticketless fans away.
"I think the fact that nine people could die at Roskilde really shocked people," he said.
"That is a highly-efficient, well-organised established Scandinavian festival that's been going for 20 years - it showed it could happen anywhere."
He said festivals had become more dangerous because of the rise of mosh pits, where hundreds of fans jump violently together.
"This sort of swell in numbers could really mean life or death," Strummer said. "We do not want people getting trampled to death in the mud here."