Geronimo the alpaca must be put down, the government has insisted, as animal rights protesters marched on Downing Street campaigning for him to be saved.

The animal, which is six years old, has twice tested positive for bovine tuberculosis and is set to be euthanised.

The Department of Food, Environment and Rural Affairs (Defra) ordered he be put down, and Geronimo’s owner, Helen Macdonald, recently lost her final appeal at the High Court in London to save her beloved creature.

Ms Macdonald, who imported Geronimo from New Zealand, believes the bovine tuberculosis tests have been returning false positives but has been refused permission to have him tested a third time.

There has been an outpouring of support from the public to save the condemned alpaca, with more than 100,000 people signing a petition calling on Boris Johnson to stop the killing.

And on Monday afternoon, protesters gathered outside Defra’s headquarters in Westminster before marching to the gates of Downing Street.

Some carried banners reading “We stand with Geronimo” and “[George] Eustice, admit you’re wrong”.

No alpacas were present at the protest due to “safety concerns”, the Born Free Foundation, which helped organise the march, said.

The protesters are demanding a different type of tuberculosis (TB) test be used to prove Geronimo’s disease status before he is put down.

But despite the mounting pressure, Downing Street insisted there will be no reprieve for Geronimo and that the case has been “looked at very carefully”.

“We know how distressing losing animals to TB is for anyone. That is why the environment secretary has looked at this extremely carefully and interrogated all the evidence,” the PM’s official spokesperson told reporters on Monday.

“The fact remains that Geronimo has sadly tested positive twice using a highly specific and reliable and validated test.

“This is something the environment secretary has looked at very carefully.”

The PM’s father Stanley Johnson has also made an intervention to save Geronimo.

But Business Secretary Kwasi Kwarteng told Kay Burley that he is “not sure where Stanley’s coming from on this”.

“My understanding is that the alpaca was tested positive for TB and the rules are that they have to be culled, because bovine TB is really, really damaging to farmers and people who make their livelihoods in agriculture and that’s why we have the policy,” he told Sky News.

Mr Kwarteng added: “I don’t think it’s a PR disaster; I think, as you say, it’s an August story. It’s obviously difficult because there’s a lot of people invested emotionally in a story of an animal, but there’s a policy and there’s no reason why the policy shouldn’t be stuck to.”

As well as alpacas, badgers have been a victim of the fight against bovine TB, with controversial mass culling employed to stop the spread since 2013.

Speaking before the protest, Dominic Dyer, from the Born Free Foundation, said: “Defra has known for many years that the TB skin test could be leading to false positive TB results in alpacas.

“However rather than allow Geronimo to be tested for TB using a more accurate Actiphage PCR blood test, Defra secretary George Eustice continues to order his death to avoid greater scrutiny over the many failures in the government’s bovine TB control policy in cattle, alpacas, and badgers.”

But writing in the Mail on Sunday, Mr Eustice said: “Farmers understand that infected animals are a risk to the remainder of their herd, so while the loss of individual animals is always a tragedy, the farming communities have worked with our government vets in this arduous but necessary endeavour.”

Ms Macdonald has threatened to film the last moments of her alpaca’s life if the killing goes ahead and broadcast it on social media.

Campaigners have joined Ms Macdonald at her farm in Wickwar, Gloucestershire, to protest against Geronimo’s impending fate.

“This is an animal injustice and everyone can relate to it and people are coming from all sectors of farming to support us,” Ms Macdonald said.

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