Excalibur, an American Staffordshire Terrior and rescue dog, was put down in October 2014 after his owner was put in isolation after contracting the virus.
A nurse at Madrid's Carlos III hospital in Madrid, Romero had been treating two Spanish missionaries who were flown home from Africa and later died.
Spanish health authorities decided to sacrifice the dog as a “precautionary measure” despite a total lack of evidence that the virus could be passed between species.
At the time Spanish health authorities were criticized for their hasty reaction.
Hundreds of people staged demonstrations calling for the dog to be saved a petition calling for the dog to be placed in quarantine instead of put down garned nearly half a million signatures in just 48 hours.
Even leading Ebola researchers supported him being kept alive.
The world authority on the virus, Eric Leroy, said at the time that "the dog does not have to be killed because it is important from a scientific point of view."
It was later discovered that the dog did not in fact have the virus.
She was only told of his death after surviving the illness and her and husband Javier Limon have fought for justice ever since.
"He was like the child we never had and we will continue asking for justice for a death that should never have happened," said the pair when legal proceedings were launched.
The pair sued the Spanish Health ministry and asked for compensation of €150,000 in “moral damages” over the killing of their pet.
But the Supreme Court has ruled that the sacrifice was lawful and that the couple are not entitled to compensation. What's more they have been ordered to pay the cost of the trial.
“There were no protocols or human resources in place to care for the animal given the potential risk to public health,” said the ruling made public this week.
Animal rights party Pacma condemned the sentence, blaming “incompetence and a total lack of empathy” by Madrid health authorities who have denied Romero and Limon compensation for “the loss of a member of their family”
The party are demanding a proper policy be put in place to deal with such situations should they arise in the future.
Romero and Limon have since adopted a new dog, an abandoned puppy called Alma.