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John Deeney, 31, from Manchester, has now taken his daughters Alyssa, six, and Darci, five, out of Partington Central Academy in protest, while he threatens to pursue a'common assault' charge.
And he says he will not return his girls until the school, and The Dean Trust which now runs it, give permission for them to wear their studs.
The row erupted when both girls - who have worn earrings since they were one - went back to class on Monday with studs in their ears.
But at the end of the day, Alyssa's were missing - and she told mum Amy Rawcliffe they had been removed by the headteacher and another teacher.
That same day, Darci told her parents she had been told off for wearing her studs.
On Tuesday, John returned with his daughters still wearing their jewellery - to be told they would have to work in isolation.
As a result, he has removed them from the school.
John, who has lodged a complaint with the school and also contacted police to see if he can press charges, said: "The first day of operating under the new trust and this happens.I checked out the website - it clearly stated no jewellery apart from inexpensive studs, which is what my girls wear.
"That a headteacher and another member of staff were standing over my daughter taking off her earrings without a call to either parent is unacceptable.
"They should not be putting their hands on my daughter and I have called police to see if this is assault." A screen grab John says he took on Monday from the policy section of the school's website - which now appears to have been removed - states "inexpensive" stud earrings can be worn.
But in another uniform section of the site, parents are warned of a jewellery ban.
And John said both his daughters had never been in trouble for wearing them before The Dean Trust took over this term.
He added: "They should not be punished because they haven't done anything wrong.
"The headteacher said the earrings could affect their behaviour and education - but as far as I've been told their behaviour is fantastic and their work is above what is expected of them." He added: "The teachers said they had changed the standard of dress code.
"Yet, I watched four members of staff, some with hooped earrings, one with a full chain in their ear. Maybe if it's so important they want to be leading by example." Tarun Kapur, chief executive of The Dean Trust, said: "As part of The Dean Trust policy on uniform, all parents were provided with advance notice that jewellery was not part of the school uniform.
"This is not at all an unusual situation in schools where we are concentrating on education rather than fashion.
"We cannot comment on individual family circumstances.
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