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The cubs, believed to be three or four months’ old, were found by members of the public on Saturday.
The pair are now in the care of officers at the National Wildlife Rescue Centre in Alloa.
Carers believe something happened to their mother and say they would not have survived alone.
Staff have named the female cub Inver and the male cub Forth.
Centre manager Steven Gray said: “Otters will stay with their mothers in the wild until they are over a year old.
“We suspect their mum was involved in a road traffic accident or some form of disturbance and didn’t return to her cubs so they have gone looking for her.
“They were seen wandering down the road in Inverkeithing on Saturday night.”
Mr Gray said the female cub was spotted by someone taking their bins out.
He said: “She was making very loud peeping noises which was probably her trying to call for her mum. The members of the public followed at a safe distance to a nearby field where they were able to monitor her until we arrived.
“The pair wouldn’t have survived on their own as they rely on mum so we are thankful the members of the public contacted us.”
The cubs will stay at the centre until they are 12 to 14 months old, as they would have stayed with their mother in the wild.
They will be taught how to feed and take care of themselves before being released when they are ready.
Mr Gray also warned anyone who may come across an otter that the animals are very agile and can have a nasty bite. They see humans as a threat and can react negatively out of fear, so it is better not to approach them.
The SSPCA has experience and the necessary safety equipment to carefully contain otters.