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The BBC reported that anyone providing short-term let (STL) accommodation must now have a licence or face a ban and fine of up to £2,500.
The changes were brought in to help to tackle problems with housing demand and anti-social behaviour, especially in STL hot spots such as Edinburgh.
But critics say the legislation is flawed and a threat to Scotland's tourism sector and wider economy.
Bed and breakfasts, homes rented out on platforms such as Airbnb and the short-term renting out of spare rooms are all affected by the changes.
Anyone who has not now applied for a licence but continues to operate can be fined and banned from applying to join the scheme for a year.
Housing Minister Paul McLennan said:
"The introduction of licensing safeguards the role short-term let accommodation plays in our economy by providing assurance to guests on safety and quality, such as gas certificate compliance and suitability of electrical equipment."
Andy Fenner, of the UK Short Term Accommodation Association, said:
"This has been a lesson on how not to pass legislation, it has been a unmitigated disaster and I don't think anyone involved could possibly say anything else.
"Our industry is growing because the demand is there, people visiting Scotland want to stay in holiday lets.
"These changes mean owners are spending large sums of money, sometimes thousands of pounds, on obtaining licences for businesses that in many cases have been there for years."