A four-year-old child has been admitted to hospital in Ayrshire with the E.coli O157 infection.
The boy who attends the Flowerbank Nursery in Kilmarnock fell ill two weeks ago.
Other youngsters at the nursery have been treated for sickness and diarrhoea but no further cases of the potentially deadly infection have been confirmed.
Children and staff who have developed symptoms since October have been asked not to attend the nursery until tested.
Dr Maida Smellie, Consultant in Public Health with NHS Ayrshire and Arran, said: "We have been working closely with the environmental health department and education services of East Ayrshire Council and with Health Protection Scotland to identify possible sources.
"Investigations are still on-going but it does not appear at this stage that the nursery is the source of the E.coli O157 infection."
She added: "Over the last three weeks, a number of children and staff at the nursery have had gastro-intestinal symptoms.
"This is consistent with a viral infection and most are recovering in two to three days."
No other children or staff have tested positive for E.coli O157 and swabs taken from surfaces at the nursery have tested negative for the infection.
The symptoms of E.Coli include feeling sick, vomiting, stomach pain, diarrhoea and fever.
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