The clip, believed to date back to 2010, shows Putin warning any betrayal of Russia would result in “choking on silver”.
The shocking video was aired on BBC Newsnight amid concerns that an assassination attempt was carried out on Russian military intelligence colonel Sergei Skripal.
The 66-year-old Russian double agent and his female companion were rushed to hospital on Sunday afternoon.
They were found unconscious at a shopping centre in Salisbury.
British counter-terror cops are aiding a probe by Wiltshire police after it was established that the pair were “exposed to an unknown substance”.
In the video clip, which has resurfaced after eight years, Putin says: "Traitors will kick the bucket. Trust me.
“These people betrayed their friends, their brothers in arms.
“Whatever they got in exchange for it, those 30 pieces of silver they were given, they will choke on them.”
Sergei Skripal was rushed to hospital following the suspected poisoning, and is said to be in a critical condition.
The Kremlin has denied suggestions Russia poisoned the double agent, with spokesman Dmitry Peskov saying Skripal's illness was "a tragic incident".
Skripal served in Russian military intelligence as a colonel until 1999, before going to work in the Foreign Ministry.
But he was arrested in Moscow in 2004, and confessed to having been recruited by British intelligence services in 1995.
Nicknamed “The Spy with the Louis Vuitton bag”, Skripal confessed to have handed over information about the network of Russian agents across Europe in exchange for around £72,500.
Skripal was charged with "high treason in the form of espionage" and was found guilty.
He was sentenced to 13 years in prison, but was later one of the four prisoners Moscow swapped in 2010 for spies in the US.
Those released during the exchange included Anna Chapman, a Russian sleeper agent operating in the USA.
The circumstances surrounding Skripal’s sudden illness echo the mysterious death of Russian spy Alexander Litvinenko, who was allegedly assassinated in a Kremlin-based attack.
The agent was killed in London in 2006 after consuming radioactive polonium-210.
He suffered for an agonising three weeks after the poison was slipped into a cup of tea.
Following his death, it was revealed the 44-year-old had been paid by MI6.
Dmitri Kovtun and Andrei Lugovoi, who both face US sanctions, are wanted by British police over the 2006 assassination of Litvinenko.
Russia has since refused to extradite Kovtun and Lugovoi so they can face trial in Britain.
Speaking to BBC Radio 4's Today programme, Alexander Goldfarb, a friend of assassinated Russian spy Alexander Litvinenko, claimed the alleged poisoning of yet another double agent is down to politics.
He said: "I have a theory, that it’s not a spy story, it’s a political story. It has to do with the elections of the president, which will happen in Russia about ten days from now.
"The major problem for Putin is the turnout. The main opponent is barred from participation, he has called for a boycott. Putin worried few people will come, they are apathetic.
"This will be used, regardless of whether Putin did it or not, as a way to show his anti-West rhetoric."
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