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Well before polls closed, one separatist leader said the region would form its own state bodies and military after the referendum, formalising a split that began with the armed takeover of state buildings in a dozen eastern towns last month.
Another said the vote would not change the region's status, but simply show that the East wanted to decide its own fate, whether in Ukraine, on its own or as part of Russia.
A near festive atmosphere at makeshift polling stations in some areas belied the potentially grave implications of the event. In others, clashes broke out between separatists and troops, over ballot papers and control of a television tower.
Zhenya Denyesh, a 20-year-old student voting early at a university building in the rebel stronghold of Slaviansk, said: "We all want to live in our own country". But asked what he thought would follow, he replied: "It will still be war."
In the southeastern port of Mariupol, scene of fierce fighting last week, there were only eight polling centres for a population of half a million. Queues grew to hundreds of metres in bright sunshine, with spirits high as one centre overflowed and ballot boxes were brought onto the street.
On the eastern outskirts, a little over an hour after polls opened, soldiers from Kiev seized what they said were falsified ballot papers, marked with Yes votes, and detained two men.
They refused to hand the men over to policemen who came to take them away, saying they did not trust them. Instead they waited for state security officers to interview and arrest them.
On the edge of Slaviansk, fighting broke out around a television tower shortly before people began making their way through barricades of felled trees, tyres and machinery for a vote Western leaders say is being orchestrated by Moscow. The Ukrainian defence ministry said one serviceman was wounded.
The West has threatened more sanctions against Russia in the key areas of energy, financial services and engineering if it continues what they regard as efforts to destabilise Ukraine.
The European Union declared the vote illegal on Sunday and may announce some modest measures as soon as Monday, limited by the bloc's reluctance to upset trade ties with Russia.
Moscow denies any role in the fighting or any ambitions to absorb the mainly Russian-speaking east, an industrial hub, into the Russian Federation following its annexation of the Black Sea peninsula of Crimea after a referendum in March.
Ukraine's Interior Ministry called the referendum a criminal farce, its ballot papers "soaked in blood". One official said that two thirds of the territory had declined to participate.
Ballot papers in the referendum in the regions of Luhansk and Donetsk, which has declared itself a "People's Republic", were printed without security provision, voter registration was patchy and there was confusion over what the vote was for. Separatists in Luhansk said only five percent had voted against.
Engineer Sergei, 33, voting in the industrial centre of Mariupol, said he would answer "Yes" to the question printed in Russian and Ukrainian on the ballot: "Do you support the act of state self-rule of the Donetsk People's Republic?"
"We're all for the independence of the Donetsk republic," he said. "It means leaving behind that fascist, pro-American government (in Kiev), which brought no one any good."
AUTONOMY, INDEPENDENCE, ANNEXATION
But in the same queue of voters, 54-year-old Irina, saw a "Yes" vote as endorsement of autonomy within Ukraine.
"I want Donetsk to have its own powers, some kind of autonomy, separate from Kiev. I'm not against a united Ukraine, but not under those people we did not choose, who seized power and are going to ruin the country," she said.
Others see the vote as a nod to absorption by Russia.
Annexation is favoured by the more prominent rebels, but the ambiguity may reflect their fears an explicit call for full "independence" might not have garnered the support they seek and could leave them in an exposed position towards Kiev.
The present government came to power when President Viktor Yanokovich was toppled in February after mass protests in Kiev.
Pro-Western activists were angered by his decision to discard a cooperation accord with the European Union in favour of closer ties with Moscow. They also accused him of corruption penetrating all areas of the Ukrainian state.
Voting is due to end in the hastily arranged referendum in 53 locations at 10 p.m. (1900 GMT) and the rebels hope to have the ballots counted by Monday afternoon, although its outcome will not be widely recognised internationally or by Kiev.
With several hours of polling to go, Russian news agencies were already reporting a turnout of more than 75 percent, although a separatist spokesman in Luhansk said troops had prevented the movement of ballot papers in several areas.
One way or another it is likely to show a large "yes" vote, and one leading separatist said Ukrainian troops would be declared illegal occupiers once results were announced.
"It is necessary to form state bodies and military authorities as soon as possible," Denis Pushilin, a leader of the self-styled Donetsk republic said, according to Interfax news agency.
Roman Lyagin, head of the rebel central electoral commission, struck a less radical tone to reporters in Donetsk.
"With the announcement of the results the status of the Donetsk region does not change in absolute terms. We do not cease to be a part of Ukraine, we do not become a part of Russia," he said, although he left those options open.
"We want only to declare to the world that we want changes ... We want to decide the fate of our region ourselves,"
Moscow has massed troops on the border and Kiev fears they may be called in as peacekeepers. Serhiy Pashinsky, head of the Ukrainian presidential administration, said a column of armoured vehicles on the Russian side of the border bore the colours of U.N. peacekeeping forces. He offered no evidence or detail.
"We warn the Kremlin that appearance of these forces on the territory of Ukraine would be assessed as military aggression and we would react as we would in the case of military aggression," he told reporters.
"INTO THE ABYSS"
Ukrainian leader Oleksander Turchinov has urged eastern political leaders to join a "Round Table" discussion on devolution of powers in Ukraine. But he says he would not negotiate with "terrorists", a formulation meant to exclude most of the more prominent rebel leaders.
Pashinsky said Ukrainian forces had "destroyed" a separatist base and checkpoints in a broad operation around Slaviansk and nearby Kramatorsk in retaliation for attacks on their posts.
"This is not a referendum. This is a desultory attempt by killers and terrorists to cover their activity," he said.
The rebels in the east and the Kremlin say the pro-European Kiev government that replaced Yanukovich lacks legitimacy.
Kiev aims to banish such questions by holding a presidential election on May 25 and the United States and European Union have threatened Russia with sweeping sanctions if it disrupts it.
Turchinov, who has ruled the referendum illegal and dismissed the allegations that the Kiev authorities are neo-fascists, said on Saturday any move to secession would be "a step into the abyss" and economic ruin.
The Metinvest company partially owned by Rinat Akhmetov, one of Ukraine's wealthiest businessmen with interests in the coal and steel industry in the east, said it was deploying a volunteer militia in Mariupol with workers from steel plants.
Akhmetov has presented himself as neutral in the conflict and Metinvest urged Kiev to refrain from sending troops into the city if his militia maintained order with police.