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Zimbabwe made 173 for seven, featuring a dashing 76 from Elton Chigumbura, in their 20 overs before holding Australia to 172 for seven.
Opener David Warner top-scored for Australia with 72 and Twenty20 captain Michael Clarke made 49 but, even though left-arm quick Mitchell Johnson had earlier taken four wickets for 23 runs, Zimbabwe denied Australia victory.
Australia needed 13 off the final over but Clarke was dimissed off the penultimate delivery before Johnson, one of several Aussies to be run out, was exited off a wide.
Brett Lee's leg-bye off the last ball was not enough for Australia.
Earlier, Chigumbura's runs came from just 35 balls with six sixes and five fours in an innings where Australia all-rounder Shane Watson's two overs went for an expensive 34 runs.
"Obviously we would have liked to have won," Johnson said.
"It's disappointing to lose in a practice match but we've just got to keep going forward, keep working hard in our training sessions and make sure we're ready for the World Cup." Clarke's side were without the rested duo of fast bowler Shaun Tait and vice-captain Cameron White.
But the duo are set to return for Thursday's second and final warm-up game against the Windward Islands.
Australia's opening match of the tournament proper is against defending champions Pakistan in St Lucia on Sunday.
In other warm-up matches, New Zealand proved too strong for minnows Ireland in a 40-run victory in Guyana.
Kiwi opener Jesse Ryder made 64 and Martin Guptill 50 as New Zealand compiled a total of 187 for five in their 20 overs.
Ireland, in reply, struggled with captain William Porterfield's 34, the top score in a total of 147 for nine. Nathan McCullum led New Zealand's attack with three for 25.
The Black Caps are in action against Sri Lanka, last year's losing finalists, and Ireland face hosts the West Indies in a 'double-bill' that will open the tournament in Guyana on Friday.
Pakistan overwhelmed the Windward Islands by 68 runs in a warm-up match in St Lucia while, in Bridgetown, Bangladesh beat Barbados by 36 runs.
