Geronimo the alpaca will be slaughtered after his owner lost a last-ditch High Court bid to save him.

The animal has twice tested positive for bovine tuberculosis and, as a result, the Department for Environment, Food and Rural Affairs (Defra) has ordered its destruction.

His owner Helen Macdonald, who imported him from New Zealand, believes the tests are returning false positives, but has been refused permission to have him tested a third time.

Earlier this month she lost her final appeal to save her beloved pet at the High Court in London and a warrant was signed for his destruction.

Ms Macdonald, who owns a farm at Wickwar, South Gloucestershire, has received an outpouring of support from the public, with more than 130,000 people signing a petition calling on Boris Johnson to halt the killing.

On Tuesday, an urgent application for a temporary injunction to halt the enforcement of the destruction order was considered by Mrs Justice Stacey at the High Court in London.

However, the judge said she would need further information from Ms Macdonald and from Government lawyers before she could make her decision, and said she would resume the hearing on Wednesday afternoon.

At court on Wednesday, the judge refused the urgent application and concluded there was “no prospect” of Ms Macdonald succeeding in her bid to reopen a previous ruling.

Ned Westaway, representing the Defra executive agency the Animal and Plant Health Agency (Apha), told the court the agency will not seek to execute the warrant on Wednesday evening, and would give Ms Macdonald the opportunity to make her own arrangements for Geronimo’s destruction.

Ms Macdonald’s lawyers told the court Geronimo first tested positive for bovine tuberculosis in September 2017 and has been in isolation since.

Catrin McGahey QC told the court that although Defra argued in previous hearings that there was a “residual risk” to other animals, the agency has also agreed Ms Macdonald’s bio-security arrangements are “impeccable”.

She said it had come to light following the publicity resulting from Ms Macdonald’s case that other animals who have been subjected to the same testing regime as Geronimo have later showed no signs of the disease after being euthanised.

Ms McGahey said: “The only issue is whether the defendants should have disclosed the fact that they had in their possession evidence that other camelids who had been subjected to repeated priming had gone on to test positive in Enferplex tests, and that there had been no sign of bovine tuberculosis on post-mortem examination.”

The barrister said the publicity had led the Daily Mail to find the owners of nine other camelids who were tested under the same regime, whose animals showed no signs of the disease after slaughter.

However, following an adjournment to allow Ms Macdonald’s lawyers to decide what evidence they wish Defra to produce and a time estimate of how long that may take, the judge refused to grant injunctions to spare Geronimo pending a further hearing and for disclosure.

Mrs Justice Stacey said that on the evidence before her, Ms Macdonald had not succeeded in showing there was any prospect of her reopening the litigation.

She said the farmer’s complaint about non-disclosure did not give rise to an arguable case, but was a “disingenuous and backdoor way of seeking a further route to appeal” when there was none left.

Last week the Government insisted all the evidence on the animal’s condition had been “looked at very carefully”.

Dr Iain McGill, a vet who has been acting as a veterinary scientific adviser to Ms Macdonald, said the decision made at the High Court on Wednesday was “incredible”.

He said: “The hard data we have shows there is no credibility whatsoever to Defra’s claims that the specificity of primed Enferplex is over 99%.

“This is utterly without merit and patently false, and they have been unable to produce any evidence whatsoever.

“Geronimo’s diagnosis is more than unsafe. It is a miscarriage of justice.”

The law firm representing Geronimo’s owner said they were “deeply disappointed” with the outcome of the hearing on Wednesday.

Olephant Solicitors said: “Further, the judge did not order the disclosure we had sought regarding nine other camelids that had previously tested positive for bovine TB after multiple priming yet showed no signs of bovine TB post-mortem.

“Unfortunately, Mrs Justice Stacey has taken at face value the statements by Defra that they, and specifically the Secretary of State, have complied with their duty of candour in bringing before the court all relevant information in applying for the warrant.

“We are surprised and concerned by that approach, given the information that has come to light in recent weeks regarding the nine other camelids.”

A Defra spokesman said: “We are sympathetic to Ms Macdonald’s situation – just as we are with everyone with animals affected by this terrible disease.

“It is for this reason that the testing results and options for Geronimo have been very carefully considered by Defra, the Animal and Plant Health Agency and its veterinary experts, as well as passing several stages of thorough legal scrutiny.

“Bovine tuberculosis is one of the greatest animal health threats we face today and causes devastation and distress for farming families and rural communities across the country while costing the taxpayer around £100m every year.

“Therefore, while nobody wants to cull animals, we need to do everything we can tackle this disease to stop it spreading and to protect the livelihoods of those affected.”