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Chancellor Rachel Reeves has said the NHS must "live within that budget" given to them - despite concerns being raised by GP surgeries over the increase to employers' national insurance contributions.
Speaking to broadcasters, Ms Reeves was repeatedly asked about the concerns raised by the likes of hospices, care homes and general practices.
Rather than being owned by the state, GPs are often operated by partnerships on contract from the health service, which are not eligible for the relief from national insurance increases that the public sector was given in the budget.
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Charities are also not eligible for the relief.
Dr David Wrigley, a GP and a deputy chair at the British Medical Association (BMA), said on social media: "Make no mistake.
"The impact of [national insurance] rise/lower salary threshold on NHS GP surgeries across the country will be monumental. Many are already on a financial tightrope due to years of neglect. We need a rapid announcement of full reimbursement."
Asked if charities, hospices and care homes would be listened to, Ms Reeves said: "We have to make a difficult sets of decisions at the budget on Wednesday to secure our public finances and also to put our public services on a firm footing.
"We announced £22.6bn worth of additional funding for our NHS over two years, as well as £3.1bn to invest in capital equipment, scanners, diagnostic equipment, new facilities in hospitals across the UK."
Pushed again, Ms Reeves said: "Well, the NHS budget has increased by £22.6bn and the NHS needs now to live within that budget.
"But it is the most generous settlement for decades and a half outside of COVID for the National Health Service and the NHS have confirmed that within that they will be able to deliver the 40,000 additional appointments every single week to bring those waiting lists down."
She added later on that the social care budget was given an extra £600m.
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Ms Reeves announced on Wednesday that employer national insurance contributions would increase from 13.8% to 15% from April 2025.
The payments will start when an employee earns £5,000 per tax year, down from the current £9,100.
Charlie King, the deputy director of external affairs for Hospice UK, said the Dorothy House hospice in Wiltshire would be hit with a £420,000 bill from the government's changes.
The hospice's most recent accounts showed it spent £14.6m on staffing in the most recent period, employing 335 staff.
Following the concerns being raised by the sector, Downing Street was asked by journalists if there would be relief.
"There is a general process whereby departments, the Department of Health for example, confirm their funding for general practices," a spokesperson said.
They added: "I think that's part of the annual GP contract process. I believe that will take place later in the year."
Other charities have explained their problems too.
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Michael Goldstein, the president of the United Synagogue, said: "If the government doesn't reverse its decision, it is vulnerable people across the country who will inevitably pay the price."
A statement from the synagogue said: "The United Synagogue employs more than 800 people across its communities, nurseries, cemeteries, kashrut and support departments and is now facing an additional tax bill of £500,000."