Russian gas supplies to Austria, Slovakia and the Czech Republic have now been halted, adding to the list of EU states left without Russian fuel in freezing mid-winter temperatures.
The EU has described the shutdown as unacceptable.
There looks to be little sign of a swift resolution of a pricing dispute that has slashed Russian gas supplies to the West. In some countries, this has forced authorities to take emergency measures with dwindling fuel reserves.
In Kiev, Ukrainian state energy firm Naftogaz said gas had stopped flowing through the last compressor station still functioning on Ukraine's western border.
Czech importer RWE Transgas said the supply in the main transit pipeline from Russia to the Czech Republic was halted at midnight.
Austria's OMV energy group, which earlier reported a dramatic fall in Russian supplies, said flows had now stopped completely.
Slovakia also said its supplies were halted.
Russia, the world's largest natural gas supplier, has accused Ukraine of stealing gas bound for Europe, while Kiev denies siphoning off gas and blames Moscow for engineering the crisis.
Ukrainian President Viktor Yushchenko has demanded that Russia resume shipments to Europe ‘immediately,’ in a letter to Russian President Dmitry Medvedev and European Commission chief José Manuel Barroso.
On Monday, Russian Prime Minister Vladimir Putin ordered energy giant Gazprom to reduce gas shipments via Ukraine by amounts equivalent to what Moscow has accused Kiev of stealing.
Talks between the two sides are due to resume tomorrow as the EU has become increasing concerned about the crisis, which began 1 January when Russia cut gas supplies to Ukraine's domestic market.
Some 80% of Russian gas exports to the EU pass through Ukraine.
